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NFC Smartphone-Based Electrochemical Microfluidic Device Integrated with Nanobody Recognition for C-Reactive Protein
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ACS Sensors

Cite this: ACS Sens. 2024, 9, 6, 3066–3074
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https://doi.org/10.1021/acssensors.4c00249
Published June 15, 2024

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Abstract

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Point-of-care testing (POCT) devices play a crucial role as tools for disease diagnostics, and the integration of biorecognition elements with electronic components into these devices widens their functionalities and facilitates the development of complex quantitative assays. Unfortunately, biosensors that exploit large conventional IgG antibodies to capture relevant biomarkers are often limited in terms of sensitivity, selectivity, and storage stability, considerably restricting the use of POCT in real-world applications. Therefore, we used nanobodies as they are more suitable for fabricating electrochemical biosensors with near-field communication (NFC) technology. Moreover, a flow-through microfluidic device was implemented in this system for the detection of C-reactive protein (CRP), an inflammation biomarker, and a model analyte. The resulting sensors not only have high sensitivity and portability but also retain automated sequential flow properties through capillary transport without the need for an external pump. We also compared the accuracy of CRP quantitative analyses between commercial PalmSens4 and NFC-based potentiostats. Furthermore, the sensor reliability was evaluated using three biological samples (artificial serum, plasma, and whole blood without any pretreatment). This platform will streamline the development of POCT devices by combining operational simplicity, low cost, fast analysis, and portability.

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Point-of-care testing (POCT) plays a crucial role in modern healthcare delivery, offering rapid and convenient diagnostic solutions at or near the patient’s location. Its primary advantage is the ability to provide real-time analysis, enabling healthcare providers to make immediate treatment decisions and improve patient outcomes. (1,2) With the growing demand for personalized and timely healthcare, as evidenced during the COVID-19 pandemic, the role of POCT continues to expand, contributing to more efficient and effective healthcare. (3−6)
POCT integrated with capillary-driven microfluidic devices has gained widespread attention over the past decade. (7−9) Traditional microfluidic devices usually require an external pump to drive fluid flow throughout the system. (10,11) In contrast, capillary forces can be induced by the surface tension of the solution to drive the flow and devices exploiting capillary forces can operate in the absence of an external pump. (12,13) As a result, POCT systems based on capillary-driven microfluidics have been implemented in various applications, such as the detection of heavy metals, pesticides, bacteria, viruses, biomarkers, and biomolecules. (14−18) The most popular examples include pregnancy and COVID-19 test kits. (19,20) These devices provide rapid results (typically within 15 min), require only a single drop of the running buffer for one-step analysis, and are inexpensive, user-friendly, and portable, but the flow control throughout the device must be accurate. (21,22) With the aim of simplifying device operability and improving its performance, we proposed a solution based on the lamination of multiple layers of transparent PET film and double-sided adhesive (DSA) tape. (23) This sensor facilitates automated fluid flow for washing the excess of targeted analytes and their detection by means of binders specific for C-reactive protein (CRP), but it can be adapted to accommodate other capture elements specific for CRP or, potentially, any other (soluble) biomarkers.
CRP is a biomarker that has been used for a long time to monitor systemic inflammation, infection, and more recently several other human pathologies. (24,25) Normal CRP levels typically fall within the range of 1–3 μg mL–1, while high CRP levels (20–400 μg mL–1) are associated with inflammation, infectious diseases, cardiovascular disease (CVDs), malignant tumors, autoimmune disease, and depression. (25−28) Although anti-CRP IgG antibodies have been traditionally used for CRP detection, their high production costs, heterogeneity after functionalization, and reliance on human or animal sources in the production process represent critical challenges. (29,30) Consequently, alternative capture elements, including antibody fragments, peptides, aptamers, polymers, and bacteriophages, have been proposed. (23,31−34) In the present work, we employed nanobodies previously isolated by phage display technology (35) because they are small recombinant proteins, inexpensive to produce, and simple to engineer adopting basic molecular biology techniques. The small size of nanobodies potentially allows for a higher binding density on the electrode surface, enhancing sensor sensitivity compared to anti-CRP IgG antibodies. (35)
In recent years, near-field communication (NFC) technology has become widespread in the field of electrochemical sensors, enhancing their functionality and ease of use since enables wireless communication and data transfer at close proximity, simplifying sensor setup, calibration, and data retrieval. (36,37) This technology allows seamless data exchange between sensors and mobile devices, providing users with an effective way to collect and analyze electrochemical data in real time. Herein, we present a smartphone-controlled NFC potentiostat integrated with a flow-through electrochemical microfluidic device via wireless communication and with the data-display conversion on Android smartphones (Figure 1a). We also compared this configuration with the performance offered by a standard potentiostat (PalmSens4) and by conventional ELISA for the evaluation CRP levels in artificial serum, plasma, and whole blood.

Figure 1

Figure 1. (a) Schematic illustration of the developed sensor obtained by combining the microfluidic device with a smartphone-based potentiostat. (b) Overall step-by-step modification on the screen-printed graphene electrodes (SPGEs). (c) Procedure for CRP detection using chronocoulometry (CC) measurement.

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Experimental Section

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The details of materials, reagents, and equipment are presented in Supporting Information, Section 1.

Preparation of Anti-CRP Nanobodies

The procedure relative to nanobody isolation and characterization was presented in detail in a previous report. (35) The best clone recovered after panning (E12) was subcloned into a pET14-derived expression vector for the production of the nanobody fused to 6xHis and to SpyTag in Escherichia coli. (31) Subsequently, the construct was transformed into BL21 (DE3) SOX cells for cytoplasmic expression and purified by metal affinity chromatography, as described previously. (38) Nanobody concentration was quantified by the Bradford method, and its quality was evaluated through SDS-PAGE and gel filtration techniques.

Electrochemical Microfluidic Device Fabrication

The details of the electrochemical microfluidic device construction, function, and testing were presented previously. (23) Briefly, the microfluidic pattern was created by using AutoCAD software. Next, a transparent PET film (Xerox) and double-sided adhesive tape (DSA, 467MP, 3M) were laser-cut using a laser cutting method (Laser engraver GCC LaserPro, C180II) to create the flow channels, which were then integrated into a sandwich-layer configuration. The fast-flow channel had a height of 350 μm, and the delayed channel had a height of 200 μm.
The microfluidic system has an opening for loading the sample onto the electrode. After an incubation time, buffer is added to a second inlet. The buffer solution is divided into two channels: the fast-flow channel and the delayed channel. Through capillary action, the buffer flows through the fast-flow channel and reaches the electrode after ca. 4 s and washes away the sample solution together with nonbonded analyte and interfering species. The buffer flowing through the delayed channel picks up a redox molecule (K3Fe(CN)6) and arrives at the electrode after a short delay (ca. 16 s). The buffer flow continued for several minutes, during which measurements are made.
A three-electrode system consisting of a working electrode (WE, 3 mm in diameter), counter electrode (CE), and reference electrode (RE) was used to perform the electrochemical analysis. Screen-printed graphene electrodes (SPGE) were fabricated by using an in-house screen-printing method with a conductive carbon-graphene ink (Sun Chemical Company, Milan, Italy). A transparent film served as the substrate to construct SPGEs. After printing, the carbon-graphene ink was dried for 1 h at 60 °C. Then, a silver/silver chloride (Ag/AgCl, Sun Chemical Company, Milan, Italy) ink was painted on the conductive pads of the RE and dried for 1 h at 60 °C. The obtained SPGE electrode was kept under dark and dry conditions when not in use to prevent the oxidization of the Ag/AgCl. The device design and integration of the microfluidic system with a smartphone are shown in Figure 1a.

Electrode Modification

In this study, anti-CRP nanobodies were anchored to the WE through the formation of covalent bonds. First, anodic pretreatment was performed on the SPGE, wherein a constant potential of 1.5 V versus Ag/AgCl was maintained for 120 s. This process generated hydroxyl groups (−OH) on the WE surface, as described in detail previously. (23) Then, the electrode was rinsed with DI water and treated with a mixed solution of 2.1 M LiCl and 40 mM NaIO4 (5 μL) to convert the surface functional groups from hydroxyl (−OH) to aldehyde (−CHO) groups. The modified electrode was allowed to incubate in the dark for 15 min before being washed with DI water. Successively, the modified electrode was functionalized with either 1 or 10 μg mL–1 of anti-CRP nanobodies for 1 h at room temperature (RT) and further washed using phosphate-buffered saline (PBS, pH 7.4). The covalent binding of the nanobody on the modified electrode was obtained by means of a Schiff base reaction, resulting in the formation of an imine bond (C═N). To ensure the stability of the covalent bond, 1 mg mL–1 solution of NaBH3CN was applied to the activated electrode for 15 min, followed by PBS washing. Subsequently, 3 mg mL–1 of casein was added (30 min at RT) to block unsaturated residues and avoid nonspecific interactions. After a final PBS washing step, the ready-to-use SPGEs were stored in a freezer at −20 °C. An overview of the overall immobilization procedure is presented in Figure 1b.

Electrochemical Detection of CRP

Electrochemical measurements were conducted using a PalmSens4 potentiostat/impedance analyzer (PalmSens BV, Netherlands), controlled by PStrace software version 5.9. To prevent convection effects that can influence the electrochemical current response, (39) chronocoulometry (CC) was preferred as the method for CRP quantification. The following CC parameters were selected: t-equilibrium of 3 s, applied potential of 0.0 V vs Ag/AgCl, t-interval of 0.1 s, analysis time of 200 s, whereas the current response was measured from 16 to 180 s. For CRP detection, 4 μL of CRP solution with concentrations ranging from 0.01 ng mL–1 to 100 μg mL–1 were introduced in the sample inlet. Subsequently, following the completion of the antigen-nanobody binding reaction, 150 μL of PBS was introduced in the buffer inlet. The chronoamperometric signal was consistently recorded until the peak signal was completed. The detection principle and procedure for CRP detection using the CC measurement are shown in Figure 1c.
The NFC potentiostat used in this study was the SIC4341 (Potentiometric sensor interface chip with NFC type2) from Silicon Craft Technology PLC., Thailand. This potentiostat was integrated with a Redmi Note 10S smartphone (Xiaomi) running Android operating system. Detailed technical information and the diagram of the printed circuit board (PCB) and the actual experimental setup can be found in Table S1 and Figure S1, respectively. To control the NFC potentiostat and electrochemical parameters, perform real-time data acquisition, process data, and present electrochemical results, we used the Chemister application (NFC eco for cyclic voltammetry (CV) and chronoamperometry). The complete operative scheme relative to the combination NFC potentiostat-Android smartphone is presented in Figure S2. The parameter setting of the NFC potentiostat was identical to that used in a PalmSens4 potentiostat. Raw data were exported as a text file, and subsequent data analysis, including plotting using Microsoft Excel and the evaluation of peak height and integrated peak area, was performed using Origin Pro.

CRP Detection in Biological Samples

Three types of samples were examined, including artificial serum (provided by Sigma-Aldrich, Warsaw, Poland), whole blood samples obtained from anonymous donors at a blood center in Warsaw, Poland, and blood plasma derived from the same whole blood samples (for details on the preparation process, see the previous report (23)). Artificial serum was diluted to 1 mg mL–1. The prepared artificial serum and plasma samples were subsequently spiked with varying CRP concentrations ranging from 10 ng mL–1 to 100 μg mL–1. Blood samples were used without any preparation process. The recovery efficacy of spiked CRP in artificial serum, plasma, and whole blood was calculated to assess the accuracy of the detection process.

Results and Discussion

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Electrochemical Characterization on the NFC and Traditional Potentiostats

Electrochemical detection using a smartphone was carried out with the NFC potentiostat, a compact device with the size of a credit card, controlled by an Android system. The potentiostat integrated into the SIC4341 microchip serves essential functions, acting as a controller for the potential waveforms, as well as a real-time data collector suitable for several electrochemical applications. This system operates as a potentiostat when connected to an NFC-enabled smartphone with the Chemister application installed.
Initially, we examined the electrochemical performance of the NFC potentiostat in comparison with the standard lab potentiostat (PalmSens4). Cyclic voltammetry (CV) was performed using 0.5 mM [Fe(CN6)]3– and 0.5 mM [Fe(CN6)]4– in 0.1 M KNO3 to study the electroanalytical functionality of the bare SPGE on both the conventional and NFC potentiostats at the following conditions: scanned potential from −0.4 to 0.6 V vs Ag/AgCl, scan rate of 25 mV s–1, potential step of 10 mV, and a time step of 200 ms. Figure 2a shows the characteristic voltammograms obtained from both potentiostats and evidences their high similarity.

Figure 2

Figure 2. (a) CVs of 5 mM Fe(CN6)3– and 5 mM Fe(CN6)3– in 0.1 M KNO3 at a scan rate of 25 mV s–1 obtained from PalmSens4, used as a positive control, and the new NFC potentiostat. (b) Electrochemical impedance spectroscopy (EIS) measurement and (c) CV measurements obtained at different steps of the electrode and after incubation with CRP in a static system using 5 mM Fe(CN6)3– and 5 mM Fe(CN6)3– containing 0.1 M KNO3 at a scan rate of 100 mV s–1, using nanobodies as immune-capture elements. All of the Nyquist plots were fitted with the Randles circuit (inset). (d) Representation of the CC measurements obtained with PalmSens4 and NFC potentiostat using nanobody-based electrochemical biosensor in the presence of CRP. (e) Linear regression comparing the average ΔQ via NFC and PalmSens4 potentiostats achieved at various CRP concentrations using CC.

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Characterization of the CRP Nanobody-Modified Electrode Surface

The immobilization efficiency of anti-CRP nanobodies is a crucial step for achieving a high antigen binding specificity. To achieve this, the hydroxyl functional groups of the oxidized electrode were first modified to incorporate aldehyde groups through an oxidation reaction. Anti-CRP nanobodies were subsequently immobilized on the oxidized electrode through an imine bond (C═N). To validate the process and assess the nanobody binding capacity for the CRP target, two analytical techniques were employed: electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). These techniques make it possible to discriminate small variations at the interface between electrode and electrolyte, as well as to assess the electron transfer efficiency of the redox couple ([Fe(CN6)]3–/4–). The EIS Nyquist plot was fitted by using the Randles equivalent circuit, as shown in Figure 2b. The bare SPGE (dashed line) showed low electron transfer resistance (or high charge transfer resistance, denoted as high Rct), indicating lower conductivity compared with the anodized electrode (blue line). However, the introduction of the anti-CRP nanobodies (green line) onto the electrode surface induced a noticeable increase in the Rct value. This observed increase strongly supports the successful immobilization of the immunocapture nanobody reagent E12. After the biosensor was coated with casein (yellow line), the subsequent addition of CRP (red line) triggered a significant Rct increment. These results suggest that the immunocomplex formed between anti-CRP nanobodies and CRP affected the electron transfer of the redox solution at the electrode interface. EIS results are consistent with the CV results, as shown in Figure 2c. Specifically, the current response was progressively reduced from Ipa = 93.7 ± 3.2 μA of the anodized electrode (blue line) to Ipa = 45.8 ± 4.6 μA of the CRP signal (red line) at each successive modification step, indicating the corresponding interference of electron transfer. Both the EIS and CV results indicated the successful nanobody immobilization on the electrode surface and the nanobody’s capacity to capture CRP.
CC was also employed to quantify CRP by using the NFC potentiostat. As presented in Figure 2d, CRP quantification was obtained by measuring the peak area or the change in charge (ΔQ). The ΔQ values obtained from NFC and a conventional potentiostat using various CRP concentrations were plotted (Figure 2e) and resulted in good agreement (R2 = 0.9982) between the two potentiostats, highlighting the potential of the NFC potentiostat as a POC diagnostic tool.
Optimal analytical conditions (see SI, Section 2, and Figure S3) with the PalmsSens4 potentiostat were identified using 1 μg mL–1 of nanobodies (Figure S3a), 1.5 V vs Ag/AgCl of anodization potential, 120 s of anodization time, 25 mM concentration of [K3Fe(CN)6], and 40 min of incubation time (Figure S3b). Specifically, nanobody concentrations higher than 1 μg mL–1 introduced steric hindrance, leading to a reduced electrochemical charge response.

CRP Detection Using Sequential Flow-Through Microfluidic Device

A comparison of analytical performances between the results obtained from PalmSens4 and the NFC potentiostat was conducted using a flow-through microfluidic device. Assay parameters were optimized to achieve the highest efficiency in terms of differentiated charge (ΔQ = ΔQCRP – ΔQcontrol). The analytical performance was initially examined at varying CRP concentrations with PalmSens4. In Figure 3a,3b it is evident that ΔQ increased as the concentration of CRP increased within the range between 0.01 and 500 ng mL–1. The ΔQ value exhibited a linear relationship with the logarithmic CRP concentration, with a correlation coefficient (R2) of 0.9941 (Figure 3a, inset) and a limit of detection (LOD) of 7.6 pg mL–1 (LOD = 3SDblank/slope), respectively. In its optimized form, the sensor detected accurately both very low CRP amounts and concentrations up to 500 ng mL–1. However, the upper limit is below the expected range of normal CRP levels in blood in the absence of inflammation. The long incubation time (40 min) contrasts the necessity for rapid POC diagnosis. Therefore, we tried to compensate for a shorter binding time (10 min) with an increased anti-CRP nanobody surface coverage (10 μg mL–1), despite the preliminary data indicating that high nanobody density could negatively affect the electrochemical current response. The data reported in Figure 3c show a linear relationship between ΔQ and the logarithm of CRP concentration in the range of 0.01–100 μg mL–1 (R2 = 0.9953), with an LOD (3SDblank/slope) of 1.18 ng mL–1 when PalmSens4 was used. Such promising results convinced us to apply the same conditions to the NFC potentiostat. As shown in Figure 3d, the linearity data were similar to those obtained with PalmSens4 and the LOD (1.79 ng mL–1) was just slightly higher. Summarizing, higher nanobody surface density leads to a significantly extended linear range at the cost of the sensitivity of the device to very low concentrations, therefore offering a suitable compromise between analytical accuracy in the physiologically relevant concentration range and analysis time.

Figure 3

Figure 3. (a) Quantitative calibration plot illustrating the relationship between the change in charge (ΔQ) and CRP concentrations and (b) its corresponding chronoamperograms using PalmSens4 Potentiostat. (c) Calibration plot between ΔQ calculated using PalmSens4 and CRP concentrations performed at high anti-CRP nanobody concentrations (10 μg mL–1) and shorter (10 min) incubation time. (d) Same as above but using the NFC potentiostat. (e) Selectivity analysis of the diagnostic device in the presence of different proteins (interleukin-6 (IL-6), fibrinogen, myoglobin, bovine serum albumin (BSA), human serum albumin (HSA)), alone or mixed together with CRP. The error bars represent the standard deviation calculated from three replicated measurements (n = 3).

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Next, we evaluated the selectivity of the developed sensor for CRP using a sample in which the biomarker was mixed with equimolar amounts of common interferents, including interleukin-6 (IL-6), fibrinogen, myoglobin, bovine serum albumin (BSA), and human serum albumin (HSA). As demonstrated in Figure 3e, CRP (100 ng mL–1) was specifically detected in the mixed sample, whereas interferents induced negligible signals. The results indicate that the biosensor possesses a high specificity toward CRP conferred by the anti-CRP nanobodies.
The result reproducibility was assessed by comparing data collected from ten independently prepared electrochemical sensors. The standard deviation (RSD) value of 8.9% (Figure S4) is within the acceptable range, according to the Association of Official Analytical Chemists (AOAC) guidelines (40) and confirmed the reproducibility of the proposed diagnostic approach, from the biosensor fabrication to the signal measurement.
The storage stability of the biosensor was thereafter investigated by comparing three conditions selected because they are normally implemented in commercial manufacturing storage settings: (i) desiccator at room temperature (RT, 20 ± 2 °C), (ii) humid box at RT, and (iii) freezer at −20 °C. As shown in Figure 4, the biosensor maintained its performance at over 80% in the first 4 weeks under all storage conditions and, when kept in the freezer, the activity reached 86.3% even after 8 weeks (Figure 4c). These results demonstrate the superior storage stability of our device over the conventional diagnostic platforms and suggest its suitability for real-world applications characterized by challenging conditions.

Figure 4

Figure 4. Storage stability of CRP biosensors under different conditions: (a) RT in a desiccator, (b) RT in a closed humid box, and (c) freezer (−20 °C), respectively. All measurements were calculated from three replicates (n = 3).

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Subsequently, the performance of our biosensor was compared to those of other devices designed for CRP quantification (Table S2). While its sensitivity is lower than that of some previously proposed systems, the LOD is still sufficient to detect CRP in the biologically relevant range spanning from ng mL–1 to μg mL–1. Remarkably, our device is inexpensive (less than 0.2 € per single biosensor, see Table S3 for details), rapid (complete analysis within 15 min), and portable. This makes it faster and more cost-effective than both ELISA and other electrochemical anti-CRP platforms, most of which require between 1 and 5 h.

Clinical Samples Analysis

We finally evaluated the capacity of our biosensor to quantify CRP in clinical samples. Three conditions were considered, namely, artificial serum, plasma, and whole blood samples. Initially, artificial serum samples were spiked with CRP concentrations ranging from 10 to 500 ng mL–1. The calculated values for ΔQ and the efficiency of the proposed system were then reported as percentages of detected CRP in comparison to the theoretical concentrations. As shown in Table S4, the recovery values ranged from 91.4 to 108%, similar to the results achieved using the PalmSens4.
CRP levels in plasma samples obtained from anonymous healthy blood donors were evaluated using both the PalmSens4 potentiostat and our NFC potentiostat. The results were further compared to those obtained by ELISA (Table 1). The paired t test conducted on the experimental results revealed no significant difference at a 95% confidence level. Consequently, the proposed biosensor can provide accurate CRP determination in real biological samples.
Table 1. CRP Concentration in Plasma Samples Evaluated by Different Methods
sampleELISA value (μg mL–1)adetected value NFC (μg mL–1)detected value PalmSens4 (μg mL–1)
11.62 ± 7.31.65 ± 2.81.73 ± 7.6
20.38 ± 3.30.40 ± 2.70.42 ± 1.5
30.73 ± 2.30.74 ± 0.80.74 ± 6.2
a

It should be noted that the results were investigated using the same samples as those reported in (23); therefore, we employed the same standard ELISA values.

Finally, the proposed method was applied to the detection of CRP in whole human blood obtained from three anonymous donors. Original blood samples contained CRP amounts in the range between 0.55 and 4.20 μg mL–1 and were further spiked with different concentrations (from 0 to 25 μg mL–1) of CRP. The results of this analysis are summarized in Table 2. The recovery was found to be within the range of 82.4–120%. The errors, which were measured as percentage relative error and relative standard error (RSD), were all less than 20% for all of the tested samples. Additionally, the feasibility of this biosensor was also evaluated with an additional ten blood samples using only the PalmSens4 potentiostat, and the detailed results can be found in Table S5. Altogether, the experimental results showed that the NFC-based system integrated with the flow-through microfluidic device can correctly quantify CRP in clinically relevant biological samples without the need for pretreatment procedures and could therefore be used for the assessment of inflammation, infections caused by bacteria or viruses, and the risk of heart disease.
Table 2. CRP Detection in Whole Blood Samples
sample nospiked value (μg mL–1)detected value (μg mL–1) NFC x̅ ± SDrecovery (%)detected value (μg mL–1) PalmSens4 x̅ ± SDrecovery (%)
104.20 4.06 
0.54.78 ± 0.31164.60 ± 1.0108
58.96 ± 1.695.19.13 ± 1.5102
2526.52 ± 0.989.328.47 ± 2.797.6
202.05 2.35 
0.52.46 ± 0.782.42.90 ± 1.1111
56.97 ± 0.998.47.82 ± 1.0110
2524.98 ± 1.491.731.19 ± 0.4115
300.55 0.77 
0.51.14 ± 3.01201.25 ± 1.195.8
55.54 ± 3.399.95.36 ± 3.391.9
2525.85 ± 4.310122.23 ± 1.285.9

Conclusions

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We have successfully developed a portable electrochemical biosensor that integrates an NFC potentiostat with a sequential flow-through microfluidic device and exploits nanobodies for the capture and quantification of CRP, providing a reliable and inexpensive diagnostic solution. Our device offers user-friendly operation, delivering the test results within 15 min at a cost of under 0.2 € per device. It has a wide linear range of detection (10 ng mL–1–100 μg mL–1) and an elevated LOD of 7.6 pg mL–1, and demonstrated high specificity for CRP, even in the presence of other proteins commonly found in serum samples. Its reliability was confirmed by the precise detection of CRP in artificial serum, plasma, and whole blood samples, eliminating the need for sample pretreatment steps. Importantly, this configuration can be potentially applied to any soluble biomarker by simply exchanging the recognition element used to capture the antigens. Thus, it offers an alternative and economically accessible method for the detection of any biomarker, particularly in settings where advanced clinical equipment is lacking.

Data Availability

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Data used for this article are available at the RepOD repository. (41)

Supporting Information

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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acssensors.4c00249.

  • Additional experimental details, scheme of the NFC potentiostat setup, cost breakdown of device, and comparison of literature CRP sensors (PDF)

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Author Information

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  • Corresponding Authors
  • Authors
    • Katarzyna Szot-Karpińska - Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, Warsaw 01-224, Poland;  Orcidhttps://orcid.org/0000-0003-3582-941X
    • Joanna Niedziółka-Jönsson - Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, Warsaw 01-224, Poland
    • Ario de Marco - Laboratory for Environmental and Life Sciences, University of Nova Gorica, Vipavska cesta 13, 5000 Nova Gorica, Slovenia;  Orcidhttps://orcid.org/0000-0001-7729-819X
  • Author Contributions

    The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

  • Notes
    The authors declare no competing financial interest.

Acknowledgments

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This research was supported by the National Centre for Research and Development (NCBR) through the EEA and Norway Grants (Project number: NOR/POLNOR/UPTURN/0060/2019) and by the CRP 20/026 grant offered by ICGEB. K.S.-K. thanks the National Science Centre Poland via a SONATA 13 grant UMO-2017/26/D/ST5/00980. The authors thank Silicon Craft Technology PLC. (Bangkok, Thailand) for providing them with an NFC potentiostat (SIC4341, Potentiometric sensor interface chip with NFC type2). S.B. thanks Dr. Kingkan Pungjunun and the team from Silicon Craft Technology PLC., and Dr. Abdulhadee Yakoh from Chulalongkorn University for their valuable explanation and suggestions regarding the NFC potentiostat.

References

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This article references 41 other publications.

  1. 1
    da Silva, E. T. S. G.; Souto, D. E. P.; Barragan, J. T. C.; de F. Giarola, J.; de Moraes, A. C. M.; Kubota, L. T. Electrochemical Biosensors in Point-of-Care Devices: Recent Advances and Future Trends. ChemElectroChem 2017, 4 (4), 778– 794,  DOI: 10.1002/celc.201600758
    Google Scholar
    1
    Electrochemical Biosensors in Point-of-Care Devices: Recent Advances and Future Trends
    da Silva, Everson T. S. G.; Souto, Denio E. P.; Barragan, Jose T. C.; Giarola, Juliana de F.; de Moraes, Ana C. M.; Kubota, Lauro T.
    ChemElectroChem (2017), 4 (4), 778-794CODEN: CHEMRA; ISSN:2196-0216. (Wiley-VCH Verlag GmbH & Co. KGaA)
    The use of biosensors in point-of-care (POC) testing devices has attracted considerable attention in the past few years, mainly because of their high specificity, portability, and relatively low cost. Coupling these devices with miniaturized electrochem. transducers has shown great potential toward simple, rapid, and cost-effective anal. that can be performed in the field, esp. for healthcare, environmental monitoring, and food quality control. For this reason, the no. of publications in this field has grown exponentially over the past decade, making it a trending topic in current research. Although great improvement has been achieved in the field of electrochem. biosensing, there are still some challenges to overcome, esp. concerning the improvement of sensing materials and miniaturization. In this Review, we summarize some of the most recent advances achieved in POC electrochem. biosensor applications, focusing on new materials and modifiers for biorecognition developed to improve sensitivity, specificity, stability, and response time.
  2. 2
    Madhurantakam, S.; Muthukumar, S.; Prasad, S. Emerging Electrochemical Biosensing Trends for Rapid Diagnosis of COVID-19 Biomarkers as Point-of-Care Platforms: A Critical Review. ACS Omega 2022, 7 (15), 12467– 12473,  DOI: 10.1021/acsomega.2c00638
    Google Scholar
    2
    Emerging Electrochemical Biosensing Trends for Rapid Diagnosis of COVID-19 Biomarkers as Point-of-Care Platforms: A Critical Review
    Madhurantakam, Sasya; Muthukumar, Sriram; Prasad, Shalini
    ACS Omega (2022), 7 (15), 12467-12473CODEN: ACSODF; ISSN:2470-1343. (American Chemical Society)
    A review. Rapid diagnosis is a crit. aspect assocd. with controlling the spread of COVID-19. Electrochem. sensor platforms are ideally suited for rapid and highly sensitive detection of biomols. This review focuses on state-of-the-art of COVID-19 biomarker detection by utilizing electrochem. biosensing platforms. Point-of-care (POC) sensing is 1 of the most promising and emerging fields in detecting and quantifying health biomarkers. Electrochem. biosensors play a major role in the development of point-of-care devices because of their high sensitivity, specificity, and ability for rapid anal. Integration of electrochem. with point-of-care technologies in the context of COVID-19 diagnosis and screening has facilitated in convenient operation, miniaturization, and portability. Identification of potential biomarkers in disease diagnosis is crucial for patient monitoring concerning SARS-CoV-2. In this review, we will discuss the choice of biomarkers in addn. to the various types of electrochem. sensors that have been developed to meet the needs of rapid detection and disease severity anal.
  3. 3
    Johnston, M.; Ates, H. C.; Glatz, R. T.; Mohsenin, H.; Schmachtenberg, R.; Göppert, N.; Huzly, D.; Urban, G. A.; Weber, W.; Dincer, C. Multiplexed Biosensor for Point-of-Care COVID-19 Monitoring: CRISPR-Powered Unamplified RNA Diagnostics and Protein-Based Therapeutic Drug Management. Mater. Today 2022, 61, 129– 138,  DOI: 10.1016/j.mattod.2022.11.001
    Google Scholar
    3
    Multiplexed biosensor for point-of-care COVID-19 monitoring: CRISPR-powered unamplified RNA diagnostics and protein-based therapeutic drug managemeant
    Johnston, Midori; Ceren Ates, H.; Glatz, Regina T.; Mohsenin, Hasti; Schmachtenberg, Rosanne; Goeppert, Nathalie; Huzly, Daniela; Urban, Gerald A.; Weber, Wilfried; Dincer, Can
    Materials Today (Oxford, United Kingdom) (2022), 61 (), 129-138CODEN: MTOUAN; ISSN:1369-7021. (Elsevier Ltd.)
    In late 2019 SARS-CoV-2 rapidly spread to become a global pandemic, therefore, measures to attenuate chains of infection, such as high-throughput screenings and isolation of carriers were taken. Prerequisite for a reasonable and democratic implementation of such measures, however, is the availability of sufficient testing opportunities (beyond reverse transcription PCR, the current gold std.). We, therefore, propose an electrochem., microfluidic multiplexed polymer-based biosensor in combination with CRISPR/Cas-powered assays for low-cost and accessible point-of-care nucleic acid testing. In this study, we simultaneously screen for and identify SARS-CoV-2 infections (Omicron-variant) in clin. specimens (Sample-to-result time: ∼30 min), employing LbuCas13a, while bypassing reverse transcription as well as target amplification of the viral RNA (LODs of 2,000 and 7,520 copies/μl for the E and RdRP genes, resp., and 50 copies/mL for combined targets), both of which are necessary for detection via PCR and other isothermal methods. In addn., we demonstrate the feasibility of combining synthetic biol.-driven assays based on different classes of biomols., in this case protein-based ss-lactam antibiotic detection, on the same device. The programmability of the effector and multiplexing capacity (up to six analytes) of our platform, in combination with a miniaturized measurement setup, including a credit card sized near field communication (NFC) potentiostat and a microperistaltic pump, provide a promising on-site tool for identifying individuals infected with variants of concern and monitoring their disease progression alongside other potential biomarkers or medication clearance.
  4. 4
    Broughton, J. P.; Deng, X.; Yu, G.; Fasching, C. L.; Servellita, V.; Singh, J.; Miao, X.; Streithorst, J. A.; Granados, A.; Sotomayor-Gonzalez, A.; Zorn, K.; Gopez, A.; Hsu, E.; Gu, W.; Miller, S.; Pan, C.-Y.; Guevara, H.; Wadford, D. A.; Chen, J. S.; Chiu, C. Y. CRISPR–Cas12-Based Detection of SARS-CoV-2. Nat. Biotechnol. 2020, 38 (7), 870– 874,  DOI: 10.1038/s41587-020-0513-4
    Google Scholar
    4
    CRISPR-Cas12-based detection of SARS-CoV-2
    Broughton, James P.; Deng, Xianding; Yu, Guixia; Fasching, Clare L.; Servellita, Venice; Singh, Jasmeet; Miao, Xin; Streithorst, Jessica A.; Granados, Andrea; Sotomayor-Gonzalez, Alicia; Zorn, Kelsey; Gopez, Allan; Hsu, Elaine; Gu, Wei; Miller, Steve; Pan, Chao-Yang; Guevara, Hugo; Wadford, Debra A.; Chen, Janice S.; Chiu, Charles Y.
    Nature Biotechnology (2020), 38 (7), 870-874CODEN: NABIF9; ISSN:1087-0156. (Nature Research)
    Abstr.: An outbreak of betacoronavirus severe acute respiratory syndrome (SARS)-CoV-2 began in Wuhan, China in Dec. 2019. COVID-19, the disease assocd. with SARS-CoV-2 infection, rapidly spread to produce a global pandemic. We report development of a rapid (<40 min), easy-to-implement and accurate CRISPR-Cas12-based lateral flow assay for detection of SARS-CoV-2 from respiratory swab RNA exts. We validated our method using contrived ref. samples and clin. samples from patients in the United States, including 36 patients with COVID-19 infection and 42 patients with other viral respiratory infections. Our CRISPR-based DETECTR assay provides a visual and faster alternative to the US Centers for Disease Control and Prevention SARS-CoV-2 real-time RT-PCR assay, with 95% pos. predictive agreement and 100% neg. predictive agreement.
  5. 5
    Yakoh, A.; Pimpitak, U.; Rengpipat, S.; Hirankarn, N.; Chailapakul, O.; Chaiyo, S. Paper-Based Electrochemical Biosensor for Diagnosing COVID-19: Detection of SARS-CoV-2 Antibodies and Antigen. Biosens. Bioelectron. 2021, 176, 112912  DOI: 10.1016/j.bios.2020.112912
    Google Scholar
    5
    Paper-based electrochemical biosensor for diagnosing COVID-19: Detection of SARS-CoV-2 antibodies and antigen
    Yakoh, Abdulhadee; Pimpitak, Umaporn; Rengpipat, Sirirat; Hirankarn, Nattiya; Chailapakul, Orawon; Chaiyo, Sudkate
    Biosensors & Bioelectronics (2021), 176 (), 112912CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
    Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is emerging as a global pandemic outbreak. To date, approx. one million deaths and over 32 million cases have been reported. This ongoing pandemic urgently requires an accurate testing device that can be used in the field in a fast manner. Serol. assays to detect antibodies have been proven to be a great complement to the std. method of reverse transcription-polymerase chain reaction (RT-PCR), particularly after the second week of infection. We have developed a specific and sensitive immunosensor for Ig detection produced against SARS-CoV-2. Unlike other lateral flow-based assays (LFAs) involving the utilization of multiple antibodies, we have reported a label-free paper-based electrochem. platform targeting SARS-CoV-2 antibodies without the specific requirement of an antibody. The presence of SARS-CoV-2 antibodies will interrupt the redox conversion of the redox indicator, resulting in a decreased current response. This electrochem. sensor was proven effective in real clin. sera from patients with satisfactory results. In addn., the proposed format was also extended to antigen detection (the spike protein of SARS-CoV-2), which presents new possibilities for diagnosing COVID-19.
  6. 6
    Kumar, A.; Parihar, A.; Panda, U.; Parihar, D. S. Microfluidics-Based Point-of-Care Testing (POCT) Devices in Dealing with Waves of COVID-19 Pandemic: The Emerging Solution. ACS Appl. Bio Mater. 2022, 5 (5), 2046– 2068,  DOI: 10.1021/acsabm.1c01320
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    6
    Microfluidics-Based Point-of-Care Testing (POCT) Devices in Dealing with Waves of COVID-19 Pandemic: The Emerging Solution
    Kumar, Avinash; Parihar, Arpana; Panda, Udwesh; Parihar, Dipesh Singh
    ACS Applied Bio Materials (2022), 5 (5), 2046-2068CODEN: AABMCB; ISSN:2576-6422. (American Chemical Society)
    A review. Recent advances in microfluidics-based point-of-care testing (POCT) technol. such as paper, array, and beads have shown promising results for diagnosing various infectious diseases. The fast and timely detection of viral infection has proven to be a crit. step for deciding the therapeutic outcome in the current COVID-19 pandemic which in turn not only enhances the patient survival rate but also reduces the disease-assocd. comorbidities. In the present scenario rapid, noninvasive detection of the virus using low cost and high throughput microfluidics-based POCT devices embraces the advantages over existing diagnostic technologies, for which need of centralized lab. facility, expensive instruments, sample pretreatment, skilled personnel required. Microfluidic-based Multiplexed POCT devices can be a boon for clin. diagnosis in developing countries that lacks a centralized health care system and resources. The microfluidic devices can be used for disease diagnosis and exploited for the development and testing of drug efficacy for disease treatment in model systems. The havoc created by the second wave of COVID-19 led several countries' governments to the back front. Lack of diagnostic kits, medical devices, and human resources created a huge demand for a technol. that can be remotely operated by single touch and data can be analyzed on phone. Recent advancements in information technol. and the use of smartphones led to a paradigm shift in the development of diagnostic devices which can be explored to deal with the current pandemic situation. This review shed light on various approaches for the development of cost-effective microfluidics POCT devices. The successfully used microfluidic devices for COVID-19 detection under clin. settings along with their pros and cons have been discussed here. Further, the integration of microfluidic devices with smartphones and wireless network systems using Internet-of-things will enable readers for manufg. advanced POCT devices for remote disease management in low resource settings.
  7. 7
    Yang, S.-M.; Lv, S.; Zhang, W.; Cui, Y. Microfluidic Point-of-Care (POC) Devices in Early Diagnosis: A Review of Opportunities and Challenges. Sensors 2022, 22 (4), 1620,  DOI: 10.3390/s22041620
    Google Scholar
    There is no corresponding record for this reference.
  8. 8
    Xie, Y.; Dai, L.; Yang, Y. Microfluidic Technology and Its Application in the Point-of-Care Testing Field. Biosens. Bioelectron.: X 2022, 10, 100109  DOI: 10.1016/j.biosx.2022.100109
    Google Scholar
    There is no corresponding record for this reference.
  9. 9
    Tang, R.; Yang, H.; Choi, J. R.; Gong, Y.; You, M.; Wen, T.; Li, A.; Li, X.; Xu, B.; Zhang, S.; Mei, Q.; Xu, F. Capillary Blood for Point-of-Care Testing. Crit. Rev. Clin. Lab. Sci. 2017, 54 (5), 294– 308,  DOI: 10.1080/10408363.2017.1343796
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    There is no corresponding record for this reference.
  10. 10
    Binsley, J. L.; Martin, E. L.; Myers, T. O.; Pagliara, S.; Ogrin, F. Y. Microfluidic Devices Powered by Integrated Elasto-Magnetic Pumps. Lab Chip 2020, 20 (22), 4285– 4295,  DOI: 10.1039/D0LC00935K
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    10
    Microfluidic devices powered by integrated elasto-magnetic pumps
    Binsley, Jacob L.; Martin, Elizabeth L.; Myers, Thomas O.; Pagliara, Stefano; Ogrin, Feodor Y.
    Lab on a Chip (2020), 20 (22), 4285-4295CODEN: LCAHAM; ISSN:1473-0189. (Royal Society of Chemistry)
    We show how an asym. elasto-magnetic system provides a novel integrated pumping soln. for lab-on-a-chip and point of care devices. This monolithic pumping soln., inspired by Purcell 's 3-link swimmer, is integrated within a simple microfluidic device, bypassing the requirement of external connections. We exptl. prove that this system can provide tuneable fluid flow with a flow rate of up to 600μL h-1. This fluid flow is achieved by actuating the pump using a weak, uniform, uniaxial, oscillating magnetic field, with field amplitudes in the range of 3-6 mT. Crucially, the fluid flow can be reversed by adjusting the driving frequency. We exptl. prove that this device can successfully operate on fluids with a range of viscosities, where pumping at higher viscosity correlates with a decreasing optimal driving frequency. The fluid flow produced by this device is understood here by examg. the non-reciprocal motion of the elasto-magnetic component. This device has the capability to replace external pumping systems with a simple, integrated, lab-on-a-chip component.
  11. 11
    Bogdanowicz, R.; Jönsson-Niedziółka, M.; Vereshchagina, E.; Dettlaff, A.; Boonkaew, S.; Pierpaoli, M.; Wittendorp, P.; Jain, S.; Tyholdt, F.; Thomas, J.; Wojcik, P. Microfluidic Devices for Photo-and Spectroelectrochemical Applications. Curr. Opin. Electrochem. 2022, 36, 101138  DOI: 10.1016/j.coelec.2022.101138
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    There is no corresponding record for this reference.
  12. 12
    Xie, Y.; Xu, X.; Wang, J.; Lin, J.; Ren, Y.; Wu, A. Latest Advances and Perspectives of Liquid Biopsy for Cancer Diagnostics Driven by Microfluidic On-Chip Assays. Lab Chip 2023, 23 (13), 2922– 2941,  DOI: 10.1039/D2LC00837H
    Google Scholar
    There is no corresponding record for this reference.
  13. 13
    Mark, D.; Haeberle, S.; Roth, G.; von Stetten, F.; Zengerle, R. Microfluidic Lab-on-a-Chip Platforms: Requirements, Characteristics and Applications. Chem. Soc. Rev. 2010, 39 (3), 1153– 1182,  DOI: 10.1039/b820557b
    Google Scholar
    13
    Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications
    Mark, Daniel; Haeberle, Stefan; Roth, Guenter; von Stetten, Felix; Zengerle, Roland
    Chemical Society Reviews (2010), 39 (3), 1153-1182CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
    A review of developments in microfluidic platforms that enable the miniaturization, integration, automation, and parallelization of (bio-)chem. assays. In contrast to isolated application-specific solns., a microfluidic platform provides a set of fluidic unit operations, which are designed for easy combination within a well-defined fabrication technol. This allows the easy, fast, and cost-efficient implementation of different application-specific (bio-)chem. processes. The review focuses on recent developments from the last decade. It is started with a brief introduction into tech. advances, major market segments and promising applications. A detailed characterization is done of different microfluidic platforms, comprising a short definition, the functional principle, microfluidic unit operations, application examples as well as strengths and limitations of every platform. The microfluidic platforms in focus are lateral flow tests, linear actuated devices, pressure driven laminar flow, microfluidic large scale integration, segmented flow microfluidics, centrifugal microfluidics, electrokinetics, electrowetting, surface acoustic waves, and dedicated systems for massively parallel anal. This review concludes with the attempt to provide a selection scheme for microfluidic platforms which is based on their characteristics according to key requirements of different applications and market segments. Applied selection criteria comprise portability, costs of instrument and disposability, sample throughput, no. of parameters per sample, reagent consumption, precision, diversity of microfluidic unit operations and the flexibility in programming different liq. handling protocols.
  14. 14
    Zhou, W.; Dou, M.; Timilsina, S. S.; Xu, F.; Li, X. Recent Innovations in Cost-Effective Polymer and Paper Hybrid Microfluidic Devices. Lab Chip 2021, 21 (14), 2658– 2683,  DOI: 10.1039/D1LC00414J
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    14
    Recent innovations in cost-effective polymer and paper hybrid microfluidic devices
    Zhou, Wan; Dou, Maowei; Timilsina, Sanjay S.; Xu, Feng; Li, XiuJun
    Lab on a Chip (2021), 21 (14), 2658-2683CODEN: LCAHAM; ISSN:1473-0189. (Royal Society of Chemistry)
    A review. Hybrid microfluidic systems that are composed of multiple different types of substrates have been recognized as a versatile and superior platform, which can draw benefits from different substrates while avoiding their limitations. This review article introduces the recent innovations of different types of low-cost hybrid microfluidic devices, particularly focusing on cost-effective polymer- and paper-based hybrid microfluidic devices. In this article, the fabrication of these hybrid microfluidic devices is briefly described and summarized. We then highlight various hybrid microfluidic systems, including polydimethylsiloxane (PDMS)-based, thermoplastic-based, paper/polymer hybrid systems, as well as other emerging hybrid systems (such as thread-based). The special benefits of using these hybrid systems have been summarized accordingly. A broad range of biol. and biomedical applications using these hybrid microfluidic devices are discussed in detail, including nucleic acid anal., protein anal., cellular anal., 3D cell culture, organ-on-a-chip, and tissue engineering. The perspective trends of hybrid microfluidic systems involving the improvement of fabrication techniques and broader applications are also discussed at the end of the review.
  15. 15
    Clark, K. M.; Schenkel, M. S.; Pittman, T. W.; Samper, I. C.; Anderson, L. B. R.; Khamcharoen, W.; Elmegerhi, S.; Perera, R.; Siangproh, W.; Kennan, A. J.; Geiss, B. J.; Dandy, D. S.; Henry, C. S. Electrochemical Capillary Driven Immunoassay for Detection of SARS-CoV-2. ACS Meas. Sci. Au 2022, 2 (6), 584– 594,  DOI: 10.1021/acsmeasuresciau.2c00037
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    15
    Electrochemical Capillary Driven Immunoassay for Detection of SARS-CoV-2
    Clark, Kaylee M.; Schenkel, Melissa S.; Pittman, Trey W.; Samper, Isabelle C.; Anderson, Loran B. R.; Khamcharoen, Wisarut; Elmegerhi, Suad; Perera, Rushika; Siangproh, Weena; Kennan, Alan J.; Geiss, Brian J.; Dandy, David S.; Henry, Charles S.
    ACS Measurement Science Au (2022), 2 (6), 584-594CODEN: AMACHV; ISSN:2694-250X. (American Chemical Society)
    The COVID-19 pandemic focused attention on a pressing need for fast, accurate, and low-cost diagnostic tests. This work presents an electrochem. capillary driven immunoassay (eCaDI) developed to detect SARS-CoV-2 nucleocapsid (N) protein. The low-cost flow device is made of polyethylene terephthalate (PET) and adhesive films. Upon addn. of a sample, reagents and washes are sequentially delivered to an integrated screen-printed carbon electrode for detection, thus automating a full sandwich immunoassay with a single end-user step. The modified electrodes are sensitive and selective for SARS-CoV-2 N protein and stable for over 7 wk. The eCaDI was tested with influenza A and Sindbis virus and proved to be selective. The eCaDI was also successfully applied to detect nine different SARS-CoV-2 variants, including Omicron.
  16. 16
    Pungjunun, K.; Praphairaksit, N.; Chailapakul, O. A Facile and Automated Microfluidic Electrochemical Platform for the In-Field Speciation Analysis of Inorganic Arsenic. Talanta 2023, 265, 124906  DOI: 10.1016/j.talanta.2023.124906
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    There is no corresponding record for this reference.
  17. 17
    Sierra, T.; Jang, I.; Noviana, E.; Crevillen, A. G.; Escarpa, A.; Henry, C. S. Pump-Free Microfluidic Device for the Electrochemical Detection of A1-Acid Glycoprotein. ACS Sens. 2021, 6 (8), 2998– 3005,  DOI: 10.1021/acssensors.1c00864
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    17
    Pump-Free Microfluidic Device for the Electrochemical Detection of α1-Acid Glycoprotein
    Sierra, Tania; Jang, Ilhoon; Noviana, Eka; Crevillen, Agustin G.; Escarpa, Alberto; Henry, Charles S.
    ACS Sensors (2021), 6 (8), 2998-3005CODEN: ASCEFJ; ISSN:2379-3694. (American Chemical Society)
    α1-Acid glycoprotein (AGP) is a glycoprotein present in serum, which is assocd. with the modulation of the immune system in response to stress or injuries, and a biomarker for inflammatory diseases and cancers. Here, we propose a pump-free microfluidic device for the electrochem. detn. of AGP. The microfluidic device utilizes capillary-driven flow and a passive mixing system to label the AGP with the Os (VI) complex (an electrochem. tag) inside the main channel, before delivering the products to the electrode surface. Furthermore, thanks to the resulting geometry, all the anal. steps can be carried out inside the device: labeling, washing, and detection by adsorptive transfer stripping square wave voltammetry. The microfluidic device exhibited a linear range from 500 to 2000 mg L-1 (R2 = 0.990) and adequate limit of detection (LOD = 231 mg L-1). Com. serum samples were analyzed to demonstrate the success of the method, yielding recoveries around 83%. Due to its simplicity, low sample consumption, low cost, short anal. time, disposability, and portability, the proposed method can serve as a point-of-care/need testing device for AGP.
  18. 18
    Jang, I.; Kang, H.; Song, S.; Dandy, D. S.; Geiss, B. J.; Henry, C. S. Flow Control in a Laminate Capillary-Driven Microfluidic Device. Analyst 2021, 146 (6), 1932– 1939,  DOI: 10.1039/D0AN02279A
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    18
    Flow control in a laminate capillary-driven microfluidic device
    Jang, Ilhoon; Kang, Hyunwoong; Song, Simon; Dandy, David S.; Geiss, Brian J.; Henry, Charles S.
    Analyst (Cambridge, United Kingdom) (2021), 146 (6), 1932-1939CODEN: ANALAO; ISSN:0003-2654. (Royal Society of Chemistry)
    Capillary-driven microfluidic devices are of significant interest for on-site anal. because they do not require external pumps and can be made from inexpensive materials. Among capillary-driven devices, those made from paper and polyester film are among the most common and have been used in a wide array of applications. However, since capillary forces are the only driving force, flow is difficult to control, and passive flow control methods such as changing the geometry must be used to accomplish various anal. applications. This study presents several new flow control methods that can be utilized in a laminate capillary-driven microfluidic device to increase available functionality. First, we introduce push and burst valve systems that can stop and start flow. These valves can stop flow for >30 min and be opened by either pressing the channel or inflowing other fluids to the valve region. Next, we propose flow control methods for Y-shaped channels that enable more functions. In one example, we demonstrate the ability to accurately control concn. to create laminar, gradient, and fully mixed flows. In a second example, flow velocity in the main channel is controlled by adjusting the length of the inlet channel. In addn., the flow velocity is const. as the inlet length increases. Finally, the flow velocity in the Y-shaped device as a function of channel height and fluid properties such as viscosity and surface tension was examd. As in previous studies on capillary-driven channels, the flow rate was affected by each parameter. The fluidic control tools presented here will enable new designs and functions for low cost point of need assays across a variety of fields.
  19. 19
    Samper, I. C.; Sánchez-Cano, A.; Khamcharoen, W.; Jang, I.; Siangproh, W.; Baldrich, E.; Geiss, B. J.; Dandy, D. S.; Henry, C. S. Electrochemical Capillary-Flow Immunoassay for Detecting Anti-SARS-CoV-2 Nucleocapsid Protein Antibodies at the Point of Care. ACS Sens. 2021, 6 (11), 4067– 4075,  DOI: 10.1021/acssensors.1c01527
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    19
    Electrochemical Capillary-Flow Immunoassay for Detecting Anti-SARS-CoV-2 Nucleocapsid Protein Antibodies at the Point of Care
    Samper, Isabelle C.; Sanchez-Cano, Ana; Khamcharoen, Wisarut; Jang, Ilhoon; Siangproh, Weena; Baldrich, Eva; Geiss, Brian J.; Dandy, David S.; Henry, Charles S.
    ACS Sensors (2021), 6 (11), 4067-4075CODEN: ASCEFJ; ISSN:2379-3694. (American Chemical Society)
    Rapid and inexpensive serol. tests for SARS-CoV-2 antibodies are needed to conduct population-level seroprevalence surveillance studies and can improve diagnostic reliability when used in combination with viral tests. We report a novel low-cost electrochem. capillary-flow device to quantify IgG antibodies targeting SARS-CoV-2 nucleocapsid proteins (anti-N antibody) down to 5 ng/mL in low-vol. (10μL) human whole blood samples in under 20 min. No sample prepn. is needed as the device integrates a blood-filtration membrane for on-board plasma extn. The device is made of stacked layers of a hydrophilic polyester and double-sided adhesive films, which create a passive microfluidic circuit that automates the steps of an ELISA. The sample and reagents are sequentially delivered to a nitrocellulose membrane that is modified with a recombinant SARS-CoV-2 nucleocapsid protein. When present in the sample, anti-N antibodies are captured on the nitrocellulose membrane and detected via chronoamperometry performed on a screen-printed carbon electrode. As a result of this quant. electrochem. readout, no result interpretation is required, making the device ideal for point-of-care (POC) use by non-trained users. Moreover, we show that the device can be coupled to a near-field communication potentiostat operated from a smartphone, confirming its true POC potential. The novelty of this work resides in the integration of sensitive electrochem. detection with capillary-flow immunoassay, providing accuracy at the point of care. This novel electrochem. capillary-flow device has the potential to aid the diagnosis of infectious diseases at the point of care.
  20. 20
    Zhu, X.; Wang, X.; Li, S.; Luo, W.; Zhang, X.; Wang, C.; Chen, Q.; Yu, S.; Tai, J.; Wang, Y. Rapid, Ultrasensitive, and Highly Specific Diagnosis of COVID-19 by CRISPR-Based Detection. ACS Sens. 2021, 6 (3), 881– 888,  DOI: 10.1021/acssensors.0c01984
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    There is no corresponding record for this reference.
  21. 21
    Andryukov, B. G. Six Decades of Lateral Flow Immunoassay: From Determining Metabolic Markers to Diagnosing COVID-19. AIMS Microbiol. 2020, 6 (3), 280– 304,  DOI: 10.3934/microbiol.2020018
    Google Scholar
    21
    Six decades of lateral flow immunoassay: from determining metabolic markers to diagnosing COVID-19
    Andryukov, Boris G.
    AIMS Microbiology (2020), 6 (3), 280-304CODEN: AMIIDH; ISSN:2471-1888. (AIMS Press)
    A review. Technologies based on lateral flow immunoassay (LFIA), known in some countries of the world as immunochromatog. tests, have been successfully used for the last six decades in diagnostics of many diseases and conditions as they allow rapid detection of mol. ligands in biosubstrates. The popularity of these diagnostic platforms is constantly increasing in healthcare facilities, particularly those facing limited budgets and time, as well as in household use for individual health monitoring. The advantages of these low-cost devices over modern lab.-based analyzers come from their availability, opportunity of rapid detection, and ease of use. The attractiveness of these portable diagnostic tools is assocd. primarily with their high anal. sensitivity and specificity, as well as with the easy visual readout of results. These qualities explain the growing popularity of LFIA in developing countries, when applied at small hospitals, in emergency situations where screening and monitoring health condition is crucially important, and as well as for self-testing of patients. These tools have passed the test of time, and now LFIA test systems are fully consistent with the world's modern concept of 'point-of-care testing', finding a wide range of applications not only in human medicine, but also in ecol., veterinary medicine, and agriculture. The extensive opportunities provided by LFIA contribute to the continuous development and improvement of this technol. and to the creation of new-generation formats. This review will highlight the modern principles of design of the most widely used formats of test-systems for clin. lab. diagnostics, summarize the main advantages and disadvantages of the method, as well as the current achievements and prospects of the LFIA technol. The latest innovations are aimed at improving the anal. performance of LFIA platforms for the diagnosis of bacterial and viral infections, including COVID-19.
  22. 22
    Perju, A.; Wongkaew, N. Integrating High-Performing Electrochemical Transducers in Lateral Flow Assay. Anal. Bioanal. Chem. 2021, 413 (22), 5535– 5549,  DOI: 10.1007/s00216-021-03301-y
    Google Scholar
    22
    Integrating high-performing electrochemical transducers in lateral flow assay
    Perju, Antonia; Wongkaew, Nongnoot
    Analytical and Bioanalytical Chemistry (2021), 413 (22), 5535-5549CODEN: ABCNBP; ISSN:1618-2642. (Springer)
    A review. Lateral flow assays (LFAs) are the best-performing and best-known point-of-care tests worldwide. Over the last decade, they have experienced an increasing interest by researchers towards improving their anal. performance while maintaining their robust assay platform. Com., visual and optical detection strategies dominate, but it is esp. the research on integrating electrochem. (EC) approaches that may have a chance to significantly improve an LFA's performance that is needed in order to detect analytes reliably at lower concns. than currently possible. In fact, EC-LFAs offer advantages in terms of quant. detn., low-cost, high sensitivity, and even simple, label-free strategies. Here, the various configurations of EC-LFAs published are summarized and critically evaluated. In short, most of them rely on applying conventional transducers, e.g., screen-printed electrode, to ensure reliability of the assay, and addnl. advances are afforded by the beneficial features of nanomaterials. It is predicted that these will be further implemented in EC-LFAs as high-performance transducers. Considering the low cost of point-of-care devices, it becomes even more important to also identify strategies that efficiently integrate nanomaterials into EC-LFAs in a high-throughput manner while maintaining their favorable anal. performance.
  23. 23
    Boonkaew, S.; Szot-Karpińska, K.; Niedziółka-Jönsson, J.; Pałys, B.; Jönsson-Niedziółka, M. Point-of-Care Testing for C-Reactive Protein in a Sequential Microfluidic Device. Sens. Actuators, B 2023, 397, 134659  DOI: 10.1016/j.snb.2023.134659
    Google Scholar
    There is no corresponding record for this reference.
  24. 24
    Sproston, N. R.; Ashworth, J. J. Role of C-Reactive Protein at Sites of Inflammation and Infection. Front. Immunol. 2018, 9, 342848,  DOI: 10.3389/fimmu.2018.00754
    Google Scholar
    There is no corresponding record for this reference.
  25. 25
    Plebani, M. Why C-Reactive Protein Is One of the Most Requested Tests in Clinical Laboratories?. Clin. Chem. Lab. Med. 2023, 61 (9), 1540– 1545,  DOI: 10.1515/cclm-2023-0086
    Google Scholar
    There is no corresponding record for this reference.
  26. 26
    Sonuç Karaboğa, M. N.; Sezgintürk, M. K. A Novel Silanization Agent Based Single Used Biosensing System: Detection of C-Reactive Protein as a Potential Alzheimer’s Disease Blood Biomarker. J. Pharm. Biomed. Anal. 2018, 154, 227– 235,  DOI: 10.1016/j.jpba.2018.03.016
    Google Scholar
    26
    A novel silanization agent based single used biosensing system: Detection of C-reactive protein as a potential Alzheimer's disease blood biomarker
    Sonuc Karaboga, Munteha Nur; Sezginturk, Mustafa Kemal
    Journal of Pharmaceutical and Biomedical Analysis (2018), 154 (), 227-235CODEN: JPBADA; ISSN:0731-7085. (Elsevier B.V.)
    This paper illustrates a new and sensitive electrochem. immunosensor for the anal. of C-reactive protein. Indium Tin Oxide (ITO) disposable sheets were modified by using 3-cyanopropyltrimethoxysilane (CPTMS) self-assembled monolayers (SAMs) for the first time for immobilizing the anti-CRP antibody via covalent interactions without the need for any crosslinking agent. Cyclic voltammetry (CV), and electrochem. impedance spectroscopy (EIS), as well as square wave voltammetry (SWV) methods were applied to characterize immobilization steps of anti-CRP and to det. the CRP concn. The optimization of the fabricated parameters and the anal. performance of the biosensor were widely evaluated. Charge transfer resistance changes were highly linear and sensitive with CRP concn. of 3.25-208 fg mL-1 range and assocd. with a limit of detection of 0.455 fg mL-1. This impedimetric biosensing system have excellent repeatability, reproducibility and reusability. Moreover, the binding characterization of CRP to anti-CRP was monitored by a single frequency impedance technique. The amt. of CRP in human serum samples were analyzed by fabricated biosensor to det. the feasibility of the biosensing system in medical purposes. We suggest that CPTMS, a new silanization agent, is ideal in biosensor applications.
  27. 27
    Kim, K.-W.; Kim, B.-M.; Moon, H.-W.; Lee, S.-H.; Kim, H.-R. Role of C-Reactive Protein in Osteoclastogenesis in Rheumatoid Arthritis. Arthritis Res. Ther. 2015, 17 (1), 41,  DOI: 10.1186/s13075-015-0563-z
    Google Scholar
    There is no corresponding record for this reference.
  28. 28
    Ali, N. Elevated Level of C-Reactive Protein May Be an Early Marker to Predict Risk for Severity of COVID-19. J. Med. Virol. 2020, 92 (11), 2409– 2411,  DOI: 10.1002/jmv.26097
    Google Scholar
    28
    Elevated level of C-reactive protein may be an early marker to predict risk for severity of COVID-19
    Ali, Nurshad
    Journal of Medical Virology (2020), 92 (11), 2409-2411CODEN: JMVIDB; ISSN:0146-6615. (Wiley-Blackwell)
    The outbreak of coronavirus disease-2019 (COVID-19) is an emerging global health threat. The healthcare workers are facing challenges in reducing the severity and mortality of COVID-19 across the world. Severe patients with COVID-19 are generally treated in the intensive care unit, while mild or non-severe patients treated in the usual isolation ward of the hospital. However, there is an emerging challenge that a small subset of mild or non-severe COVID-19 patients develops in to a severe disease course. Serum C-reactive protein (CRP) has been found as an important marker that changes significantly in severe patients with COVID-19. The elevated levels of CRP might be linked to the overprodn. of inflammatory cytokines in severe patients with COVID-19. However, CRP levels in patients with COVID-19 who may progress from non-severe to severe cases need to be further studied in large-scale multicenter studies.
  29. 29
    Bryan, T.; Luo, X.; Bueno, P. R.; Davis, J. J. An Optimised Electrochemical Biosensor for the Label-Free Detection of C-Reactive Protein in Blood. Biosens. Bioelectron. 2013, 39 (1), 94– 98,  DOI: 10.1016/j.bios.2012.06.051
    Google Scholar
    There is no corresponding record for this reference.
  30. 30
    Chames, P.; Van Regenmortel, M.; Weiss, E.; Baty, D. Therapeutic Antibodies: Successes, Limitations and Hopes for the Future. Br. J. Pharmacol. 2009, 157 (2), 220– 233,  DOI: 10.1111/j.1476-5381.2009.00190.x
    Google Scholar
    30
    Therapeutic antibodies: successes, limitations and hopes for the future
    Chames, Patrick; Van Regenmortel, Marc; Weiss, Etienne; Baty, Daniel
    British Journal of Pharmacology (2009), 157 (2), 220-233CODEN: BJPCBM; ISSN:1476-5381. (Wiley-Blackwell)
    A review. With more than 20 mols. in clin. use, monoclonal antibodies have finally come of age as therapeutics, generating a market value of $11 billion in 2004, expected to reach $26 billion by 2010. While delivering interesting results in the treatment of several major diseases including autoimmune, cardiovascular and infectious diseases, cancer and inflammation, clin. trials and research are generating a wealth of useful information, for instance about assocns. of clin. responses with Fc receptor polymorphisms and the infiltration and recruitment of effector cells into targeted tissues. Some functional limitations of therapeutic antibodies have come to light such as inadequate pharmacokinetics and tissue accessibility as well as impaired interactions with the immune system, and these deficiencies point to areas where addnl. research is needed. This review aims at giving an overview of the current state of the art and describes the most promising avenues that are being followed to create the next generation of antibody-based therapeutic agents.
  31. 31
    Oloketuyi, S.; Mazzega, E.; Zavašnik, J.; Pungjunun, K.; Kalcher, K.; Marco, A.; de Mehmeti, E. Electrochemical Immunosensor Functionalized with Nanobodies for the Detection of the Toxic Microalgae Alexandrium Minutum Using Glassy Carbon Electrode Modified with Gold Nanoparticles. Biosens. Bioelectron. 2020, 154, 112052,  DOI: 10.1016/j.bios.2020.112052
    Google Scholar
    31
    Electrochemical immunosensor functionalized with nanobodies for the detection of the toxic microalgae Alexandrium minutum using glassy carbon electrode modified with gold nanoparticles
    Oloketuyi, Sandra; Mazzega, Elisa; Zavasnik, Janez; Pungjunun, Kingkan; Kalcher, Kurt; de Marco, Ario; Mehmeti, Eda
    Biosensors & Bioelectronics (2020), 154 (), 112052CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
    In this work an electrochem. immunosensor for the toxic microalgae Alexandrium minutum (A. minutum AL9T) detection is described. A glassy carbon electrode (GCE) was modified by depositing gold nanoparticles followed by L-cysteine for obtaining a self-assembled monolayer. The SpyTagged nanobody C1, specific for the A. minutum toxic strain AL9T, was then covalently immobilized via SpyCatcher on the surface of the modified electrode and used for the selective capture of such microalgae strain. Electrochem. impedance spectroscopy (EIS) was used for the quantification of A. minutum cells present in water samples by measuring the charge-transfer resistance changes of the electrode with a hexacyanoferrate probe. Each electrode modification step was accompanied by cyclic voltammetry (CV) and SEM (SEM). The immunosensor provided highly reproducible data, was simple to fabricate at low cost, exhibited higher sensitivity than previously described alternative diagnostic methods and showed a broad linear range between 103 and 109 cells L-1 with detection limit of 3 × 103 cells L-1 of A. minutum AL9T. The immunosensor was successfully applied to quantify A. minutum AL9T in seawater and brackish water samples proving that it can be used for early detection of harmful microalgae without the necessity of pre-concn. or dialysis steps.
  32. 32
    Szot-Karpińska, K.; Kudła, P.; Orzeł, U.; Narajczyk, M.; Jönsson-Niedziółka, M.; Pałys, B.; Filipek, S.; Ebner, A.; Niedziółka-Jönsson, J. Investigation of Peptides for Molecular Recognition of C-Reactive Protein–Theoretical and Experimental Studies. Anal. Chem. 2023, 95, 14475,  DOI: 10.1021/acs.analchem.3c03127
    Google Scholar
    There is no corresponding record for this reference.
  33. 33
    Yang, H. J.; Kim, M. W.; Raju, C. V.; Cho, C. H.; Park, T. J.; Park, J. P. Highly Sensitive and Label-Free Electrochemical Detection of C-Reactive Protein on a Peptide Receptor–gold Nanoparticle–black Phosphorous Nanocomposite Modified Electrode. Biosens. Bioelectron. 2023, 234, 115382  DOI: 10.1016/j.bios.2023.115382
    Google Scholar
    33
    Highly sensitive and label-free electrochemical detection of C-reactive protein on a peptide receptor-gold nanoparticle-black phosphorous nanocomposite modified electrode
    Yang, Hyo Jeong; Kim, Min Woo; Raju, Chikkili Venkateswara; Cho, Chae Hwan; Park, Tae Jung; Park, Jong Pil
    Biosensors & Bioelectronics (2023), 234 (), 115382CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
    C-reactive protein (CRP) is a phylogenetically highly conserved plasma protein found in blood serum, and an enhanced CRP level is indicative of inflammatory conditions such as infection and cancer, among others. In this work, we developed a novel high CRP-affinity peptide-functionalized label-free electrochem. biosensor for the highly sensitive and selective detection of CRP. Throughout biopanning with random peptide libraries, high affinity peptides for CRP was successfully identified, and then a series of synthetic peptide receptor, of which C-terminus was incorporated to gold binding peptide (GBP) as an anchoring motif was covalently immobilized onto gold nanoparticle (AuNPs) tethered polydopamine (PDA)-black phosphorus (BP) (AuNPs@BP@PDA) nanocomposite electrode. Interaction between the CRP-binding peptide and CRP was confirmed via ELISA along with various physicochem. and electrochem. analyses. Under the optimized exptl. conditions, the proposed peptide-based biosensor detects CRP in the range of 0-0.036 μg/mL with a detection limit (LOD) of 0.7 ng/mL. The developed sensor effectively detects CRP in the real samples of serum and plasma of Crohns disease patients. Thus, the fabricated peptide-based biosensor has potential applications in clin. diagnosis and medical applications.
  34. 34
    Szot-Karpińska, K.; Kudła, P.; Szarota, A.; Narajczyk, M.; Marken, F.; Niedziółka-Jönsson, J. CRP-Binding Bacteriophage as a New Element of Layer-by-Layer Assembly Carbon Nanofiber Modified Electrodes. Bioelectrochemistry 2020, 136, 107629  DOI: 10.1016/j.bioelechem.2020.107629
    Google Scholar
    34
    CRP-binding bacteriophage as a new element of layer-by-layer assembly carbon nanofiber modified electrodes
    Szot-Karpinska, Katarzyna; Kudla, Patryk; Szarota, Anna; Narajczyk, Magdalena; Marken, Frank; Niedziolka-Jonsson, Joanna
    Bioelectrochemistry (2020), 136 (), 107629CODEN: BIOEFK; ISSN:1567-5394. (Elsevier B.V.)
    Recently, bacteriophage particles have started to be applied as a new biomaterial for developing sensing platforms. They can be used as both a recognition element or/and as building blocks, template/scaffold. In this paper, we studied a bacteriophage selected through phage-display technol. The chosen bacteriophage acted as a building block for creating a carbon nanofiber-based electrode and as a new receptor/binding element that recognizes C-reactive protein (CRP) - one of the markers of inflammatory processes in the human body. The binding efficiency of the selected phage towards CRP is two orders of magnitude higher than in the wild type. We demonstrate that the phage-based sensor is selective against other proteins. Finally, we show that layer-by-layer methods are suitable for deposition of neg. charged phages (wild or CRP-binding) with pos. charged carbon nanofibers for electrode surface modification. A three-layered electrode was successfully used for mol. recognition of CRP, and the mol. interactions were studied using electrochem., biol., and optical methods, including microscopic and spectroscopic anal.
  35. 35
    Oloketuyi, S.; Bernedo, R.; Christmann, A.; Borkowska, J.; Cazzaniga, G.; Schuchmann, H. W.; Niedziółka-Jönsson, J.; Szot-Karpińska, K.; Kolmar, H.; de Marco, A. Native Llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection. Biosensors 2021, 11 (12), 496,  DOI: 10.3390/bios11120496
    Google Scholar
    There is no corresponding record for this reference.
  36. 36
    Beck, J. J.; Alenicheva, V.; Rahn, K. L.; Russo, M. J.; Baldo, T. A.; Henry, C. S. Evaluating the Performance of an Inexpensive, Commercially Available, NFC-Powered and Smartphone Controlled Potentiostat for Electrochemical Sensing. Electroanalysis 2023, 35 (6), e202200552  DOI: 10.1002/elan.202200552
    Google Scholar
    There is no corresponding record for this reference.
  37. 37
    Pungjunun, K.; Yakoh, A.; Chaiyo, S.; Siangproh, W.; Praphairaksit, N.; Chailapakul, O. Smartphone-Based Electrochemical Analysis Integrated with NFC System for the Voltammetric Detection of Heavy Metals Using a Screen-Printed Graphene Electrode. Microchim. Acta 2022, 189 (5), 191,  DOI: 10.1007/s00604-022-05281-x
    Google Scholar
    There is no corresponding record for this reference.
  38. 38
    Veggiani, G.; de Marco, A. Improved Quantitative and Qualitative Production of Single-Domain Intrabodies Mediated by the Co-Expression of Erv1p Sulfhydryl Oxidase. Protein Expression Purif. 2011, 79 (1), 111– 114,  DOI: 10.1016/j.pep.2011.03.005
    Google Scholar
    There is no corresponding record for this reference.
  39. 39
    Amatore, C.; Pebay, C.; Sella, C.; Thouin, L. Mass Transport at Microband Electrodes: Transient, Quasi-Steady-State, and Convective Regimes. ChemPhysChem 2012, 13 (6), 1562– 1568,  DOI: 10.1002/cphc.201100942
    Google Scholar
    There is no corresponding record for this reference.
  40. 40
    International, A. AOAC Peer-Verified Methods Program: Manual on Policies and Procedures; Association of Official Analytical Chemists, 1993.
    Google Scholar
    There is no corresponding record for this reference.
  41. 41
    Jönsson-Niedziolka, M.; Boonkaew, S. Electrochemical Data for NFC Smartphone-Based Electrochemical Microfluidic Device Integrated with Nanobody Recognition for C-Reactive Protein, RepOD – Repository for Open Data, 2024. https://doi.org/10.18150/M8JBP9 (accessed Feb 01, 2024).
    Google Scholar
    There is no corresponding record for this reference.

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  • Abstract

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    Figure 1

    Figure 1. (a) Schematic illustration of the developed sensor obtained by combining the microfluidic device with a smartphone-based potentiostat. (b) Overall step-by-step modification on the screen-printed graphene electrodes (SPGEs). (c) Procedure for CRP detection using chronocoulometry (CC) measurement.

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    Figure 2

    Figure 2. (a) CVs of 5 mM Fe(CN6)3– and 5 mM Fe(CN6)3– in 0.1 M KNO3 at a scan rate of 25 mV s–1 obtained from PalmSens4, used as a positive control, and the new NFC potentiostat. (b) Electrochemical impedance spectroscopy (EIS) measurement and (c) CV measurements obtained at different steps of the electrode and after incubation with CRP in a static system using 5 mM Fe(CN6)3– and 5 mM Fe(CN6)3– containing 0.1 M KNO3 at a scan rate of 100 mV s–1, using nanobodies as immune-capture elements. All of the Nyquist plots were fitted with the Randles circuit (inset). (d) Representation of the CC measurements obtained with PalmSens4 and NFC potentiostat using nanobody-based electrochemical biosensor in the presence of CRP. (e) Linear regression comparing the average ΔQ via NFC and PalmSens4 potentiostats achieved at various CRP concentrations using CC.

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    Figure 3

    Figure 3. (a) Quantitative calibration plot illustrating the relationship between the change in charge (ΔQ) and CRP concentrations and (b) its corresponding chronoamperograms using PalmSens4 Potentiostat. (c) Calibration plot between ΔQ calculated using PalmSens4 and CRP concentrations performed at high anti-CRP nanobody concentrations (10 μg mL–1) and shorter (10 min) incubation time. (d) Same as above but using the NFC potentiostat. (e) Selectivity analysis of the diagnostic device in the presence of different proteins (interleukin-6 (IL-6), fibrinogen, myoglobin, bovine serum albumin (BSA), human serum albumin (HSA)), alone or mixed together with CRP. The error bars represent the standard deviation calculated from three replicated measurements (n = 3).

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    Figure 4

    Figure 4. Storage stability of CRP biosensors under different conditions: (a) RT in a desiccator, (b) RT in a closed humid box, and (c) freezer (−20 °C), respectively. All measurements were calculated from three replicates (n = 3).

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  • References

    This article references 41 other publications.

    1. 1
      da Silva, E. T. S. G.; Souto, D. E. P.; Barragan, J. T. C.; de F. Giarola, J.; de Moraes, A. C. M.; Kubota, L. T. Electrochemical Biosensors in Point-of-Care Devices: Recent Advances and Future Trends. ChemElectroChem 2017, 4 (4), 778– 794,  DOI: 10.1002/celc.201600758
      1
      Electrochemical Biosensors in Point-of-Care Devices: Recent Advances and Future Trends
      da Silva, Everson T. S. G.; Souto, Denio E. P.; Barragan, Jose T. C.; Giarola, Juliana de F.; de Moraes, Ana C. M.; Kubota, Lauro T.
      ChemElectroChem (2017), 4 (4), 778-794CODEN: CHEMRA; ISSN:2196-0216. (Wiley-VCH Verlag GmbH & Co. KGaA)
      The use of biosensors in point-of-care (POC) testing devices has attracted considerable attention in the past few years, mainly because of their high specificity, portability, and relatively low cost. Coupling these devices with miniaturized electrochem. transducers has shown great potential toward simple, rapid, and cost-effective anal. that can be performed in the field, esp. for healthcare, environmental monitoring, and food quality control. For this reason, the no. of publications in this field has grown exponentially over the past decade, making it a trending topic in current research. Although great improvement has been achieved in the field of electrochem. biosensing, there are still some challenges to overcome, esp. concerning the improvement of sensing materials and miniaturization. In this Review, we summarize some of the most recent advances achieved in POC electrochem. biosensor applications, focusing on new materials and modifiers for biorecognition developed to improve sensitivity, specificity, stability, and response time.
    2. 2
      Madhurantakam, S.; Muthukumar, S.; Prasad, S. Emerging Electrochemical Biosensing Trends for Rapid Diagnosis of COVID-19 Biomarkers as Point-of-Care Platforms: A Critical Review. ACS Omega 2022, 7 (15), 12467– 12473,  DOI: 10.1021/acsomega.2c00638
      2
      Emerging Electrochemical Biosensing Trends for Rapid Diagnosis of COVID-19 Biomarkers as Point-of-Care Platforms: A Critical Review
      Madhurantakam, Sasya; Muthukumar, Sriram; Prasad, Shalini
      ACS Omega (2022), 7 (15), 12467-12473CODEN: ACSODF; ISSN:2470-1343. (American Chemical Society)
      A review. Rapid diagnosis is a crit. aspect assocd. with controlling the spread of COVID-19. Electrochem. sensor platforms are ideally suited for rapid and highly sensitive detection of biomols. This review focuses on state-of-the-art of COVID-19 biomarker detection by utilizing electrochem. biosensing platforms. Point-of-care (POC) sensing is 1 of the most promising and emerging fields in detecting and quantifying health biomarkers. Electrochem. biosensors play a major role in the development of point-of-care devices because of their high sensitivity, specificity, and ability for rapid anal. Integration of electrochem. with point-of-care technologies in the context of COVID-19 diagnosis and screening has facilitated in convenient operation, miniaturization, and portability. Identification of potential biomarkers in disease diagnosis is crucial for patient monitoring concerning SARS-CoV-2. In this review, we will discuss the choice of biomarkers in addn. to the various types of electrochem. sensors that have been developed to meet the needs of rapid detection and disease severity anal.
    3. 3
      Johnston, M.; Ates, H. C.; Glatz, R. T.; Mohsenin, H.; Schmachtenberg, R.; Göppert, N.; Huzly, D.; Urban, G. A.; Weber, W.; Dincer, C. Multiplexed Biosensor for Point-of-Care COVID-19 Monitoring: CRISPR-Powered Unamplified RNA Diagnostics and Protein-Based Therapeutic Drug Management. Mater. Today 2022, 61, 129– 138,  DOI: 10.1016/j.mattod.2022.11.001
      3
      Multiplexed biosensor for point-of-care COVID-19 monitoring: CRISPR-powered unamplified RNA diagnostics and protein-based therapeutic drug managemeant
      Johnston, Midori; Ceren Ates, H.; Glatz, Regina T.; Mohsenin, Hasti; Schmachtenberg, Rosanne; Goeppert, Nathalie; Huzly, Daniela; Urban, Gerald A.; Weber, Wilfried; Dincer, Can
      Materials Today (Oxford, United Kingdom) (2022), 61 (), 129-138CODEN: MTOUAN; ISSN:1369-7021. (Elsevier Ltd.)
      In late 2019 SARS-CoV-2 rapidly spread to become a global pandemic, therefore, measures to attenuate chains of infection, such as high-throughput screenings and isolation of carriers were taken. Prerequisite for a reasonable and democratic implementation of such measures, however, is the availability of sufficient testing opportunities (beyond reverse transcription PCR, the current gold std.). We, therefore, propose an electrochem., microfluidic multiplexed polymer-based biosensor in combination with CRISPR/Cas-powered assays for low-cost and accessible point-of-care nucleic acid testing. In this study, we simultaneously screen for and identify SARS-CoV-2 infections (Omicron-variant) in clin. specimens (Sample-to-result time: ∼30 min), employing LbuCas13a, while bypassing reverse transcription as well as target amplification of the viral RNA (LODs of 2,000 and 7,520 copies/μl for the E and RdRP genes, resp., and 50 copies/mL for combined targets), both of which are necessary for detection via PCR and other isothermal methods. In addn., we demonstrate the feasibility of combining synthetic biol.-driven assays based on different classes of biomols., in this case protein-based ss-lactam antibiotic detection, on the same device. The programmability of the effector and multiplexing capacity (up to six analytes) of our platform, in combination with a miniaturized measurement setup, including a credit card sized near field communication (NFC) potentiostat and a microperistaltic pump, provide a promising on-site tool for identifying individuals infected with variants of concern and monitoring their disease progression alongside other potential biomarkers or medication clearance.
    4. 4
      Broughton, J. P.; Deng, X.; Yu, G.; Fasching, C. L.; Servellita, V.; Singh, J.; Miao, X.; Streithorst, J. A.; Granados, A.; Sotomayor-Gonzalez, A.; Zorn, K.; Gopez, A.; Hsu, E.; Gu, W.; Miller, S.; Pan, C.-Y.; Guevara, H.; Wadford, D. A.; Chen, J. S.; Chiu, C. Y. CRISPR–Cas12-Based Detection of SARS-CoV-2. Nat. Biotechnol. 2020, 38 (7), 870– 874,  DOI: 10.1038/s41587-020-0513-4
      4
      CRISPR-Cas12-based detection of SARS-CoV-2
      Broughton, James P.; Deng, Xianding; Yu, Guixia; Fasching, Clare L.; Servellita, Venice; Singh, Jasmeet; Miao, Xin; Streithorst, Jessica A.; Granados, Andrea; Sotomayor-Gonzalez, Alicia; Zorn, Kelsey; Gopez, Allan; Hsu, Elaine; Gu, Wei; Miller, Steve; Pan, Chao-Yang; Guevara, Hugo; Wadford, Debra A.; Chen, Janice S.; Chiu, Charles Y.
      Nature Biotechnology (2020), 38 (7), 870-874CODEN: NABIF9; ISSN:1087-0156. (Nature Research)
      Abstr.: An outbreak of betacoronavirus severe acute respiratory syndrome (SARS)-CoV-2 began in Wuhan, China in Dec. 2019. COVID-19, the disease assocd. with SARS-CoV-2 infection, rapidly spread to produce a global pandemic. We report development of a rapid (<40 min), easy-to-implement and accurate CRISPR-Cas12-based lateral flow assay for detection of SARS-CoV-2 from respiratory swab RNA exts. We validated our method using contrived ref. samples and clin. samples from patients in the United States, including 36 patients with COVID-19 infection and 42 patients with other viral respiratory infections. Our CRISPR-based DETECTR assay provides a visual and faster alternative to the US Centers for Disease Control and Prevention SARS-CoV-2 real-time RT-PCR assay, with 95% pos. predictive agreement and 100% neg. predictive agreement.
    5. 5
      Yakoh, A.; Pimpitak, U.; Rengpipat, S.; Hirankarn, N.; Chailapakul, O.; Chaiyo, S. Paper-Based Electrochemical Biosensor for Diagnosing COVID-19: Detection of SARS-CoV-2 Antibodies and Antigen. Biosens. Bioelectron. 2021, 176, 112912  DOI: 10.1016/j.bios.2020.112912
      5
      Paper-based electrochemical biosensor for diagnosing COVID-19: Detection of SARS-CoV-2 antibodies and antigen
      Yakoh, Abdulhadee; Pimpitak, Umaporn; Rengpipat, Sirirat; Hirankarn, Nattiya; Chailapakul, Orawon; Chaiyo, Sudkate
      Biosensors & Bioelectronics (2021), 176 (), 112912CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
      Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is emerging as a global pandemic outbreak. To date, approx. one million deaths and over 32 million cases have been reported. This ongoing pandemic urgently requires an accurate testing device that can be used in the field in a fast manner. Serol. assays to detect antibodies have been proven to be a great complement to the std. method of reverse transcription-polymerase chain reaction (RT-PCR), particularly after the second week of infection. We have developed a specific and sensitive immunosensor for Ig detection produced against SARS-CoV-2. Unlike other lateral flow-based assays (LFAs) involving the utilization of multiple antibodies, we have reported a label-free paper-based electrochem. platform targeting SARS-CoV-2 antibodies without the specific requirement of an antibody. The presence of SARS-CoV-2 antibodies will interrupt the redox conversion of the redox indicator, resulting in a decreased current response. This electrochem. sensor was proven effective in real clin. sera from patients with satisfactory results. In addn., the proposed format was also extended to antigen detection (the spike protein of SARS-CoV-2), which presents new possibilities for diagnosing COVID-19.
    6. 6
      Kumar, A.; Parihar, A.; Panda, U.; Parihar, D. S. Microfluidics-Based Point-of-Care Testing (POCT) Devices in Dealing with Waves of COVID-19 Pandemic: The Emerging Solution. ACS Appl. Bio Mater. 2022, 5 (5), 2046– 2068,  DOI: 10.1021/acsabm.1c01320
      6
      Microfluidics-Based Point-of-Care Testing (POCT) Devices in Dealing with Waves of COVID-19 Pandemic: The Emerging Solution
      Kumar, Avinash; Parihar, Arpana; Panda, Udwesh; Parihar, Dipesh Singh
      ACS Applied Bio Materials (2022), 5 (5), 2046-2068CODEN: AABMCB; ISSN:2576-6422. (American Chemical Society)
      A review. Recent advances in microfluidics-based point-of-care testing (POCT) technol. such as paper, array, and beads have shown promising results for diagnosing various infectious diseases. The fast and timely detection of viral infection has proven to be a crit. step for deciding the therapeutic outcome in the current COVID-19 pandemic which in turn not only enhances the patient survival rate but also reduces the disease-assocd. comorbidities. In the present scenario rapid, noninvasive detection of the virus using low cost and high throughput microfluidics-based POCT devices embraces the advantages over existing diagnostic technologies, for which need of centralized lab. facility, expensive instruments, sample pretreatment, skilled personnel required. Microfluidic-based Multiplexed POCT devices can be a boon for clin. diagnosis in developing countries that lacks a centralized health care system and resources. The microfluidic devices can be used for disease diagnosis and exploited for the development and testing of drug efficacy for disease treatment in model systems. The havoc created by the second wave of COVID-19 led several countries' governments to the back front. Lack of diagnostic kits, medical devices, and human resources created a huge demand for a technol. that can be remotely operated by single touch and data can be analyzed on phone. Recent advancements in information technol. and the use of smartphones led to a paradigm shift in the development of diagnostic devices which can be explored to deal with the current pandemic situation. This review shed light on various approaches for the development of cost-effective microfluidics POCT devices. The successfully used microfluidic devices for COVID-19 detection under clin. settings along with their pros and cons have been discussed here. Further, the integration of microfluidic devices with smartphones and wireless network systems using Internet-of-things will enable readers for manufg. advanced POCT devices for remote disease management in low resource settings.
    7. 7
      Yang, S.-M.; Lv, S.; Zhang, W.; Cui, Y. Microfluidic Point-of-Care (POC) Devices in Early Diagnosis: A Review of Opportunities and Challenges. Sensors 2022, 22 (4), 1620,  DOI: 10.3390/s22041620
      There is no corresponding record for this reference.
    8. 8
      Xie, Y.; Dai, L.; Yang, Y. Microfluidic Technology and Its Application in the Point-of-Care Testing Field. Biosens. Bioelectron.: X 2022, 10, 100109  DOI: 10.1016/j.biosx.2022.100109
      There is no corresponding record for this reference.
    9. 9
      Tang, R.; Yang, H.; Choi, J. R.; Gong, Y.; You, M.; Wen, T.; Li, A.; Li, X.; Xu, B.; Zhang, S.; Mei, Q.; Xu, F. Capillary Blood for Point-of-Care Testing. Crit. Rev. Clin. Lab. Sci. 2017, 54 (5), 294– 308,  DOI: 10.1080/10408363.2017.1343796
      There is no corresponding record for this reference.
    10. 10
      Binsley, J. L.; Martin, E. L.; Myers, T. O.; Pagliara, S.; Ogrin, F. Y. Microfluidic Devices Powered by Integrated Elasto-Magnetic Pumps. Lab Chip 2020, 20 (22), 4285– 4295,  DOI: 10.1039/D0LC00935K
      10
      Microfluidic devices powered by integrated elasto-magnetic pumps
      Binsley, Jacob L.; Martin, Elizabeth L.; Myers, Thomas O.; Pagliara, Stefano; Ogrin, Feodor Y.
      Lab on a Chip (2020), 20 (22), 4285-4295CODEN: LCAHAM; ISSN:1473-0189. (Royal Society of Chemistry)
      We show how an asym. elasto-magnetic system provides a novel integrated pumping soln. for lab-on-a-chip and point of care devices. This monolithic pumping soln., inspired by Purcell 's 3-link swimmer, is integrated within a simple microfluidic device, bypassing the requirement of external connections. We exptl. prove that this system can provide tuneable fluid flow with a flow rate of up to 600μL h-1. This fluid flow is achieved by actuating the pump using a weak, uniform, uniaxial, oscillating magnetic field, with field amplitudes in the range of 3-6 mT. Crucially, the fluid flow can be reversed by adjusting the driving frequency. We exptl. prove that this device can successfully operate on fluids with a range of viscosities, where pumping at higher viscosity correlates with a decreasing optimal driving frequency. The fluid flow produced by this device is understood here by examg. the non-reciprocal motion of the elasto-magnetic component. This device has the capability to replace external pumping systems with a simple, integrated, lab-on-a-chip component.
    11. 11
      Bogdanowicz, R.; Jönsson-Niedziółka, M.; Vereshchagina, E.; Dettlaff, A.; Boonkaew, S.; Pierpaoli, M.; Wittendorp, P.; Jain, S.; Tyholdt, F.; Thomas, J.; Wojcik, P. Microfluidic Devices for Photo-and Spectroelectrochemical Applications. Curr. Opin. Electrochem. 2022, 36, 101138  DOI: 10.1016/j.coelec.2022.101138
      There is no corresponding record for this reference.
    12. 12
      Xie, Y.; Xu, X.; Wang, J.; Lin, J.; Ren, Y.; Wu, A. Latest Advances and Perspectives of Liquid Biopsy for Cancer Diagnostics Driven by Microfluidic On-Chip Assays. Lab Chip 2023, 23 (13), 2922– 2941,  DOI: 10.1039/D2LC00837H
      There is no corresponding record for this reference.
    13. 13
      Mark, D.; Haeberle, S.; Roth, G.; von Stetten, F.; Zengerle, R. Microfluidic Lab-on-a-Chip Platforms: Requirements, Characteristics and Applications. Chem. Soc. Rev. 2010, 39 (3), 1153– 1182,  DOI: 10.1039/b820557b
      13
      Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications
      Mark, Daniel; Haeberle, Stefan; Roth, Guenter; von Stetten, Felix; Zengerle, Roland
      Chemical Society Reviews (2010), 39 (3), 1153-1182CODEN: CSRVBR; ISSN:0306-0012. (Royal Society of Chemistry)
      A review of developments in microfluidic platforms that enable the miniaturization, integration, automation, and parallelization of (bio-)chem. assays. In contrast to isolated application-specific solns., a microfluidic platform provides a set of fluidic unit operations, which are designed for easy combination within a well-defined fabrication technol. This allows the easy, fast, and cost-efficient implementation of different application-specific (bio-)chem. processes. The review focuses on recent developments from the last decade. It is started with a brief introduction into tech. advances, major market segments and promising applications. A detailed characterization is done of different microfluidic platforms, comprising a short definition, the functional principle, microfluidic unit operations, application examples as well as strengths and limitations of every platform. The microfluidic platforms in focus are lateral flow tests, linear actuated devices, pressure driven laminar flow, microfluidic large scale integration, segmented flow microfluidics, centrifugal microfluidics, electrokinetics, electrowetting, surface acoustic waves, and dedicated systems for massively parallel anal. This review concludes with the attempt to provide a selection scheme for microfluidic platforms which is based on their characteristics according to key requirements of different applications and market segments. Applied selection criteria comprise portability, costs of instrument and disposability, sample throughput, no. of parameters per sample, reagent consumption, precision, diversity of microfluidic unit operations and the flexibility in programming different liq. handling protocols.
    14. 14
      Zhou, W.; Dou, M.; Timilsina, S. S.; Xu, F.; Li, X. Recent Innovations in Cost-Effective Polymer and Paper Hybrid Microfluidic Devices. Lab Chip 2021, 21 (14), 2658– 2683,  DOI: 10.1039/D1LC00414J
      14
      Recent innovations in cost-effective polymer and paper hybrid microfluidic devices
      Zhou, Wan; Dou, Maowei; Timilsina, Sanjay S.; Xu, Feng; Li, XiuJun
      Lab on a Chip (2021), 21 (14), 2658-2683CODEN: LCAHAM; ISSN:1473-0189. (Royal Society of Chemistry)
      A review. Hybrid microfluidic systems that are composed of multiple different types of substrates have been recognized as a versatile and superior platform, which can draw benefits from different substrates while avoiding their limitations. This review article introduces the recent innovations of different types of low-cost hybrid microfluidic devices, particularly focusing on cost-effective polymer- and paper-based hybrid microfluidic devices. In this article, the fabrication of these hybrid microfluidic devices is briefly described and summarized. We then highlight various hybrid microfluidic systems, including polydimethylsiloxane (PDMS)-based, thermoplastic-based, paper/polymer hybrid systems, as well as other emerging hybrid systems (such as thread-based). The special benefits of using these hybrid systems have been summarized accordingly. A broad range of biol. and biomedical applications using these hybrid microfluidic devices are discussed in detail, including nucleic acid anal., protein anal., cellular anal., 3D cell culture, organ-on-a-chip, and tissue engineering. The perspective trends of hybrid microfluidic systems involving the improvement of fabrication techniques and broader applications are also discussed at the end of the review.
    15. 15
      Clark, K. M.; Schenkel, M. S.; Pittman, T. W.; Samper, I. C.; Anderson, L. B. R.; Khamcharoen, W.; Elmegerhi, S.; Perera, R.; Siangproh, W.; Kennan, A. J.; Geiss, B. J.; Dandy, D. S.; Henry, C. S. Electrochemical Capillary Driven Immunoassay for Detection of SARS-CoV-2. ACS Meas. Sci. Au 2022, 2 (6), 584– 594,  DOI: 10.1021/acsmeasuresciau.2c00037
      15
      Electrochemical Capillary Driven Immunoassay for Detection of SARS-CoV-2
      Clark, Kaylee M.; Schenkel, Melissa S.; Pittman, Trey W.; Samper, Isabelle C.; Anderson, Loran B. R.; Khamcharoen, Wisarut; Elmegerhi, Suad; Perera, Rushika; Siangproh, Weena; Kennan, Alan J.; Geiss, Brian J.; Dandy, David S.; Henry, Charles S.
      ACS Measurement Science Au (2022), 2 (6), 584-594CODEN: AMACHV; ISSN:2694-250X. (American Chemical Society)
      The COVID-19 pandemic focused attention on a pressing need for fast, accurate, and low-cost diagnostic tests. This work presents an electrochem. capillary driven immunoassay (eCaDI) developed to detect SARS-CoV-2 nucleocapsid (N) protein. The low-cost flow device is made of polyethylene terephthalate (PET) and adhesive films. Upon addn. of a sample, reagents and washes are sequentially delivered to an integrated screen-printed carbon electrode for detection, thus automating a full sandwich immunoassay with a single end-user step. The modified electrodes are sensitive and selective for SARS-CoV-2 N protein and stable for over 7 wk. The eCaDI was tested with influenza A and Sindbis virus and proved to be selective. The eCaDI was also successfully applied to detect nine different SARS-CoV-2 variants, including Omicron.
    16. 16
      Pungjunun, K.; Praphairaksit, N.; Chailapakul, O. A Facile and Automated Microfluidic Electrochemical Platform for the In-Field Speciation Analysis of Inorganic Arsenic. Talanta 2023, 265, 124906  DOI: 10.1016/j.talanta.2023.124906
      There is no corresponding record for this reference.
    17. 17
      Sierra, T.; Jang, I.; Noviana, E.; Crevillen, A. G.; Escarpa, A.; Henry, C. S. Pump-Free Microfluidic Device for the Electrochemical Detection of A1-Acid Glycoprotein. ACS Sens. 2021, 6 (8), 2998– 3005,  DOI: 10.1021/acssensors.1c00864
      17
      Pump-Free Microfluidic Device for the Electrochemical Detection of α1-Acid Glycoprotein
      Sierra, Tania; Jang, Ilhoon; Noviana, Eka; Crevillen, Agustin G.; Escarpa, Alberto; Henry, Charles S.
      ACS Sensors (2021), 6 (8), 2998-3005CODEN: ASCEFJ; ISSN:2379-3694. (American Chemical Society)
      α1-Acid glycoprotein (AGP) is a glycoprotein present in serum, which is assocd. with the modulation of the immune system in response to stress or injuries, and a biomarker for inflammatory diseases and cancers. Here, we propose a pump-free microfluidic device for the electrochem. detn. of AGP. The microfluidic device utilizes capillary-driven flow and a passive mixing system to label the AGP with the Os (VI) complex (an electrochem. tag) inside the main channel, before delivering the products to the electrode surface. Furthermore, thanks to the resulting geometry, all the anal. steps can be carried out inside the device: labeling, washing, and detection by adsorptive transfer stripping square wave voltammetry. The microfluidic device exhibited a linear range from 500 to 2000 mg L-1 (R2 = 0.990) and adequate limit of detection (LOD = 231 mg L-1). Com. serum samples were analyzed to demonstrate the success of the method, yielding recoveries around 83%. Due to its simplicity, low sample consumption, low cost, short anal. time, disposability, and portability, the proposed method can serve as a point-of-care/need testing device for AGP.
    18. 18
      Jang, I.; Kang, H.; Song, S.; Dandy, D. S.; Geiss, B. J.; Henry, C. S. Flow Control in a Laminate Capillary-Driven Microfluidic Device. Analyst 2021, 146 (6), 1932– 1939,  DOI: 10.1039/D0AN02279A
      18
      Flow control in a laminate capillary-driven microfluidic device
      Jang, Ilhoon; Kang, Hyunwoong; Song, Simon; Dandy, David S.; Geiss, Brian J.; Henry, Charles S.
      Analyst (Cambridge, United Kingdom) (2021), 146 (6), 1932-1939CODEN: ANALAO; ISSN:0003-2654. (Royal Society of Chemistry)
      Capillary-driven microfluidic devices are of significant interest for on-site anal. because they do not require external pumps and can be made from inexpensive materials. Among capillary-driven devices, those made from paper and polyester film are among the most common and have been used in a wide array of applications. However, since capillary forces are the only driving force, flow is difficult to control, and passive flow control methods such as changing the geometry must be used to accomplish various anal. applications. This study presents several new flow control methods that can be utilized in a laminate capillary-driven microfluidic device to increase available functionality. First, we introduce push and burst valve systems that can stop and start flow. These valves can stop flow for >30 min and be opened by either pressing the channel or inflowing other fluids to the valve region. Next, we propose flow control methods for Y-shaped channels that enable more functions. In one example, we demonstrate the ability to accurately control concn. to create laminar, gradient, and fully mixed flows. In a second example, flow velocity in the main channel is controlled by adjusting the length of the inlet channel. In addn., the flow velocity is const. as the inlet length increases. Finally, the flow velocity in the Y-shaped device as a function of channel height and fluid properties such as viscosity and surface tension was examd. As in previous studies on capillary-driven channels, the flow rate was affected by each parameter. The fluidic control tools presented here will enable new designs and functions for low cost point of need assays across a variety of fields.
    19. 19
      Samper, I. C.; Sánchez-Cano, A.; Khamcharoen, W.; Jang, I.; Siangproh, W.; Baldrich, E.; Geiss, B. J.; Dandy, D. S.; Henry, C. S. Electrochemical Capillary-Flow Immunoassay for Detecting Anti-SARS-CoV-2 Nucleocapsid Protein Antibodies at the Point of Care. ACS Sens. 2021, 6 (11), 4067– 4075,  DOI: 10.1021/acssensors.1c01527
      19
      Electrochemical Capillary-Flow Immunoassay for Detecting Anti-SARS-CoV-2 Nucleocapsid Protein Antibodies at the Point of Care
      Samper, Isabelle C.; Sanchez-Cano, Ana; Khamcharoen, Wisarut; Jang, Ilhoon; Siangproh, Weena; Baldrich, Eva; Geiss, Brian J.; Dandy, David S.; Henry, Charles S.
      ACS Sensors (2021), 6 (11), 4067-4075CODEN: ASCEFJ; ISSN:2379-3694. (American Chemical Society)
      Rapid and inexpensive serol. tests for SARS-CoV-2 antibodies are needed to conduct population-level seroprevalence surveillance studies and can improve diagnostic reliability when used in combination with viral tests. We report a novel low-cost electrochem. capillary-flow device to quantify IgG antibodies targeting SARS-CoV-2 nucleocapsid proteins (anti-N antibody) down to 5 ng/mL in low-vol. (10μL) human whole blood samples in under 20 min. No sample prepn. is needed as the device integrates a blood-filtration membrane for on-board plasma extn. The device is made of stacked layers of a hydrophilic polyester and double-sided adhesive films, which create a passive microfluidic circuit that automates the steps of an ELISA. The sample and reagents are sequentially delivered to a nitrocellulose membrane that is modified with a recombinant SARS-CoV-2 nucleocapsid protein. When present in the sample, anti-N antibodies are captured on the nitrocellulose membrane and detected via chronoamperometry performed on a screen-printed carbon electrode. As a result of this quant. electrochem. readout, no result interpretation is required, making the device ideal for point-of-care (POC) use by non-trained users. Moreover, we show that the device can be coupled to a near-field communication potentiostat operated from a smartphone, confirming its true POC potential. The novelty of this work resides in the integration of sensitive electrochem. detection with capillary-flow immunoassay, providing accuracy at the point of care. This novel electrochem. capillary-flow device has the potential to aid the diagnosis of infectious diseases at the point of care.
    20. 20
      Zhu, X.; Wang, X.; Li, S.; Luo, W.; Zhang, X.; Wang, C.; Chen, Q.; Yu, S.; Tai, J.; Wang, Y. Rapid, Ultrasensitive, and Highly Specific Diagnosis of COVID-19 by CRISPR-Based Detection. ACS Sens. 2021, 6 (3), 881– 888,  DOI: 10.1021/acssensors.0c01984
      There is no corresponding record for this reference.
    21. 21
      Andryukov, B. G. Six Decades of Lateral Flow Immunoassay: From Determining Metabolic Markers to Diagnosing COVID-19. AIMS Microbiol. 2020, 6 (3), 280– 304,  DOI: 10.3934/microbiol.2020018
      21
      Six decades of lateral flow immunoassay: from determining metabolic markers to diagnosing COVID-19
      Andryukov, Boris G.
      AIMS Microbiology (2020), 6 (3), 280-304CODEN: AMIIDH; ISSN:2471-1888. (AIMS Press)
      A review. Technologies based on lateral flow immunoassay (LFIA), known in some countries of the world as immunochromatog. tests, have been successfully used for the last six decades in diagnostics of many diseases and conditions as they allow rapid detection of mol. ligands in biosubstrates. The popularity of these diagnostic platforms is constantly increasing in healthcare facilities, particularly those facing limited budgets and time, as well as in household use for individual health monitoring. The advantages of these low-cost devices over modern lab.-based analyzers come from their availability, opportunity of rapid detection, and ease of use. The attractiveness of these portable diagnostic tools is assocd. primarily with their high anal. sensitivity and specificity, as well as with the easy visual readout of results. These qualities explain the growing popularity of LFIA in developing countries, when applied at small hospitals, in emergency situations where screening and monitoring health condition is crucially important, and as well as for self-testing of patients. These tools have passed the test of time, and now LFIA test systems are fully consistent with the world's modern concept of 'point-of-care testing', finding a wide range of applications not only in human medicine, but also in ecol., veterinary medicine, and agriculture. The extensive opportunities provided by LFIA contribute to the continuous development and improvement of this technol. and to the creation of new-generation formats. This review will highlight the modern principles of design of the most widely used formats of test-systems for clin. lab. diagnostics, summarize the main advantages and disadvantages of the method, as well as the current achievements and prospects of the LFIA technol. The latest innovations are aimed at improving the anal. performance of LFIA platforms for the diagnosis of bacterial and viral infections, including COVID-19.
    22. 22
      Perju, A.; Wongkaew, N. Integrating High-Performing Electrochemical Transducers in Lateral Flow Assay. Anal. Bioanal. Chem. 2021, 413 (22), 5535– 5549,  DOI: 10.1007/s00216-021-03301-y
      22
      Integrating high-performing electrochemical transducers in lateral flow assay
      Perju, Antonia; Wongkaew, Nongnoot
      Analytical and Bioanalytical Chemistry (2021), 413 (22), 5535-5549CODEN: ABCNBP; ISSN:1618-2642. (Springer)
      A review. Lateral flow assays (LFAs) are the best-performing and best-known point-of-care tests worldwide. Over the last decade, they have experienced an increasing interest by researchers towards improving their anal. performance while maintaining their robust assay platform. Com., visual and optical detection strategies dominate, but it is esp. the research on integrating electrochem. (EC) approaches that may have a chance to significantly improve an LFA's performance that is needed in order to detect analytes reliably at lower concns. than currently possible. In fact, EC-LFAs offer advantages in terms of quant. detn., low-cost, high sensitivity, and even simple, label-free strategies. Here, the various configurations of EC-LFAs published are summarized and critically evaluated. In short, most of them rely on applying conventional transducers, e.g., screen-printed electrode, to ensure reliability of the assay, and addnl. advances are afforded by the beneficial features of nanomaterials. It is predicted that these will be further implemented in EC-LFAs as high-performance transducers. Considering the low cost of point-of-care devices, it becomes even more important to also identify strategies that efficiently integrate nanomaterials into EC-LFAs in a high-throughput manner while maintaining their favorable anal. performance.
    23. 23
      Boonkaew, S.; Szot-Karpińska, K.; Niedziółka-Jönsson, J.; Pałys, B.; Jönsson-Niedziółka, M. Point-of-Care Testing for C-Reactive Protein in a Sequential Microfluidic Device. Sens. Actuators, B 2023, 397, 134659  DOI: 10.1016/j.snb.2023.134659
      There is no corresponding record for this reference.
    24. 24
      Sproston, N. R.; Ashworth, J. J. Role of C-Reactive Protein at Sites of Inflammation and Infection. Front. Immunol. 2018, 9, 342848,  DOI: 10.3389/fimmu.2018.00754
      There is no corresponding record for this reference.
    25. 25
      Plebani, M. Why C-Reactive Protein Is One of the Most Requested Tests in Clinical Laboratories?. Clin. Chem. Lab. Med. 2023, 61 (9), 1540– 1545,  DOI: 10.1515/cclm-2023-0086
      There is no corresponding record for this reference.
    26. 26
      Sonuç Karaboğa, M. N.; Sezgintürk, M. K. A Novel Silanization Agent Based Single Used Biosensing System: Detection of C-Reactive Protein as a Potential Alzheimer’s Disease Blood Biomarker. J. Pharm. Biomed. Anal. 2018, 154, 227– 235,  DOI: 10.1016/j.jpba.2018.03.016
      26
      A novel silanization agent based single used biosensing system: Detection of C-reactive protein as a potential Alzheimer's disease blood biomarker
      Sonuc Karaboga, Munteha Nur; Sezginturk, Mustafa Kemal
      Journal of Pharmaceutical and Biomedical Analysis (2018), 154 (), 227-235CODEN: JPBADA; ISSN:0731-7085. (Elsevier B.V.)
      This paper illustrates a new and sensitive electrochem. immunosensor for the anal. of C-reactive protein. Indium Tin Oxide (ITO) disposable sheets were modified by using 3-cyanopropyltrimethoxysilane (CPTMS) self-assembled monolayers (SAMs) for the first time for immobilizing the anti-CRP antibody via covalent interactions without the need for any crosslinking agent. Cyclic voltammetry (CV), and electrochem. impedance spectroscopy (EIS), as well as square wave voltammetry (SWV) methods were applied to characterize immobilization steps of anti-CRP and to det. the CRP concn. The optimization of the fabricated parameters and the anal. performance of the biosensor were widely evaluated. Charge transfer resistance changes were highly linear and sensitive with CRP concn. of 3.25-208 fg mL-1 range and assocd. with a limit of detection of 0.455 fg mL-1. This impedimetric biosensing system have excellent repeatability, reproducibility and reusability. Moreover, the binding characterization of CRP to anti-CRP was monitored by a single frequency impedance technique. The amt. of CRP in human serum samples were analyzed by fabricated biosensor to det. the feasibility of the biosensing system in medical purposes. We suggest that CPTMS, a new silanization agent, is ideal in biosensor applications.
    27. 27
      Kim, K.-W.; Kim, B.-M.; Moon, H.-W.; Lee, S.-H.; Kim, H.-R. Role of C-Reactive Protein in Osteoclastogenesis in Rheumatoid Arthritis. Arthritis Res. Ther. 2015, 17 (1), 41,  DOI: 10.1186/s13075-015-0563-z
      There is no corresponding record for this reference.
    28. 28
      Ali, N. Elevated Level of C-Reactive Protein May Be an Early Marker to Predict Risk for Severity of COVID-19. J. Med. Virol. 2020, 92 (11), 2409– 2411,  DOI: 10.1002/jmv.26097
      28
      Elevated level of C-reactive protein may be an early marker to predict risk for severity of COVID-19
      Ali, Nurshad
      Journal of Medical Virology (2020), 92 (11), 2409-2411CODEN: JMVIDB; ISSN:0146-6615. (Wiley-Blackwell)
      The outbreak of coronavirus disease-2019 (COVID-19) is an emerging global health threat. The healthcare workers are facing challenges in reducing the severity and mortality of COVID-19 across the world. Severe patients with COVID-19 are generally treated in the intensive care unit, while mild or non-severe patients treated in the usual isolation ward of the hospital. However, there is an emerging challenge that a small subset of mild or non-severe COVID-19 patients develops in to a severe disease course. Serum C-reactive protein (CRP) has been found as an important marker that changes significantly in severe patients with COVID-19. The elevated levels of CRP might be linked to the overprodn. of inflammatory cytokines in severe patients with COVID-19. However, CRP levels in patients with COVID-19 who may progress from non-severe to severe cases need to be further studied in large-scale multicenter studies.
    29. 29
      Bryan, T.; Luo, X.; Bueno, P. R.; Davis, J. J. An Optimised Electrochemical Biosensor for the Label-Free Detection of C-Reactive Protein in Blood. Biosens. Bioelectron. 2013, 39 (1), 94– 98,  DOI: 10.1016/j.bios.2012.06.051
      There is no corresponding record for this reference.
    30. 30
      Chames, P.; Van Regenmortel, M.; Weiss, E.; Baty, D. Therapeutic Antibodies: Successes, Limitations and Hopes for the Future. Br. J. Pharmacol. 2009, 157 (2), 220– 233,  DOI: 10.1111/j.1476-5381.2009.00190.x
      30
      Therapeutic antibodies: successes, limitations and hopes for the future
      Chames, Patrick; Van Regenmortel, Marc; Weiss, Etienne; Baty, Daniel
      British Journal of Pharmacology (2009), 157 (2), 220-233CODEN: BJPCBM; ISSN:1476-5381. (Wiley-Blackwell)
      A review. With more than 20 mols. in clin. use, monoclonal antibodies have finally come of age as therapeutics, generating a market value of $11 billion in 2004, expected to reach $26 billion by 2010. While delivering interesting results in the treatment of several major diseases including autoimmune, cardiovascular and infectious diseases, cancer and inflammation, clin. trials and research are generating a wealth of useful information, for instance about assocns. of clin. responses with Fc receptor polymorphisms and the infiltration and recruitment of effector cells into targeted tissues. Some functional limitations of therapeutic antibodies have come to light such as inadequate pharmacokinetics and tissue accessibility as well as impaired interactions with the immune system, and these deficiencies point to areas where addnl. research is needed. This review aims at giving an overview of the current state of the art and describes the most promising avenues that are being followed to create the next generation of antibody-based therapeutic agents.
    31. 31
      Oloketuyi, S.; Mazzega, E.; Zavašnik, J.; Pungjunun, K.; Kalcher, K.; Marco, A.; de Mehmeti, E. Electrochemical Immunosensor Functionalized with Nanobodies for the Detection of the Toxic Microalgae Alexandrium Minutum Using Glassy Carbon Electrode Modified with Gold Nanoparticles. Biosens. Bioelectron. 2020, 154, 112052,  DOI: 10.1016/j.bios.2020.112052
      31
      Electrochemical immunosensor functionalized with nanobodies for the detection of the toxic microalgae Alexandrium minutum using glassy carbon electrode modified with gold nanoparticles
      Oloketuyi, Sandra; Mazzega, Elisa; Zavasnik, Janez; Pungjunun, Kingkan; Kalcher, Kurt; de Marco, Ario; Mehmeti, Eda
      Biosensors & Bioelectronics (2020), 154 (), 112052CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
      In this work an electrochem. immunosensor for the toxic microalgae Alexandrium minutum (A. minutum AL9T) detection is described. A glassy carbon electrode (GCE) was modified by depositing gold nanoparticles followed by L-cysteine for obtaining a self-assembled monolayer. The SpyTagged nanobody C1, specific for the A. minutum toxic strain AL9T, was then covalently immobilized via SpyCatcher on the surface of the modified electrode and used for the selective capture of such microalgae strain. Electrochem. impedance spectroscopy (EIS) was used for the quantification of A. minutum cells present in water samples by measuring the charge-transfer resistance changes of the electrode with a hexacyanoferrate probe. Each electrode modification step was accompanied by cyclic voltammetry (CV) and SEM (SEM). The immunosensor provided highly reproducible data, was simple to fabricate at low cost, exhibited higher sensitivity than previously described alternative diagnostic methods and showed a broad linear range between 103 and 109 cells L-1 with detection limit of 3 × 103 cells L-1 of A. minutum AL9T. The immunosensor was successfully applied to quantify A. minutum AL9T in seawater and brackish water samples proving that it can be used for early detection of harmful microalgae without the necessity of pre-concn. or dialysis steps.
    32. 32
      Szot-Karpińska, K.; Kudła, P.; Orzeł, U.; Narajczyk, M.; Jönsson-Niedziółka, M.; Pałys, B.; Filipek, S.; Ebner, A.; Niedziółka-Jönsson, J. Investigation of Peptides for Molecular Recognition of C-Reactive Protein–Theoretical and Experimental Studies. Anal. Chem. 2023, 95, 14475,  DOI: 10.1021/acs.analchem.3c03127
      There is no corresponding record for this reference.
    33. 33
      Yang, H. J.; Kim, M. W.; Raju, C. V.; Cho, C. H.; Park, T. J.; Park, J. P. Highly Sensitive and Label-Free Electrochemical Detection of C-Reactive Protein on a Peptide Receptor–gold Nanoparticle–black Phosphorous Nanocomposite Modified Electrode. Biosens. Bioelectron. 2023, 234, 115382  DOI: 10.1016/j.bios.2023.115382
      33
      Highly sensitive and label-free electrochemical detection of C-reactive protein on a peptide receptor-gold nanoparticle-black phosphorous nanocomposite modified electrode
      Yang, Hyo Jeong; Kim, Min Woo; Raju, Chikkili Venkateswara; Cho, Chae Hwan; Park, Tae Jung; Park, Jong Pil
      Biosensors & Bioelectronics (2023), 234 (), 115382CODEN: BBIOE4; ISSN:0956-5663. (Elsevier B.V.)
      C-reactive protein (CRP) is a phylogenetically highly conserved plasma protein found in blood serum, and an enhanced CRP level is indicative of inflammatory conditions such as infection and cancer, among others. In this work, we developed a novel high CRP-affinity peptide-functionalized label-free electrochem. biosensor for the highly sensitive and selective detection of CRP. Throughout biopanning with random peptide libraries, high affinity peptides for CRP was successfully identified, and then a series of synthetic peptide receptor, of which C-terminus was incorporated to gold binding peptide (GBP) as an anchoring motif was covalently immobilized onto gold nanoparticle (AuNPs) tethered polydopamine (PDA)-black phosphorus (BP) (AuNPs@BP@PDA) nanocomposite electrode. Interaction between the CRP-binding peptide and CRP was confirmed via ELISA along with various physicochem. and electrochem. analyses. Under the optimized exptl. conditions, the proposed peptide-based biosensor detects CRP in the range of 0-0.036 μg/mL with a detection limit (LOD) of 0.7 ng/mL. The developed sensor effectively detects CRP in the real samples of serum and plasma of Crohns disease patients. Thus, the fabricated peptide-based biosensor has potential applications in clin. diagnosis and medical applications.
    34. 34
      Szot-Karpińska, K.; Kudła, P.; Szarota, A.; Narajczyk, M.; Marken, F.; Niedziółka-Jönsson, J. CRP-Binding Bacteriophage as a New Element of Layer-by-Layer Assembly Carbon Nanofiber Modified Electrodes. Bioelectrochemistry 2020, 136, 107629  DOI: 10.1016/j.bioelechem.2020.107629
      34
      CRP-binding bacteriophage as a new element of layer-by-layer assembly carbon nanofiber modified electrodes
      Szot-Karpinska, Katarzyna; Kudla, Patryk; Szarota, Anna; Narajczyk, Magdalena; Marken, Frank; Niedziolka-Jonsson, Joanna
      Bioelectrochemistry (2020), 136 (), 107629CODEN: BIOEFK; ISSN:1567-5394. (Elsevier B.V.)
      Recently, bacteriophage particles have started to be applied as a new biomaterial for developing sensing platforms. They can be used as both a recognition element or/and as building blocks, template/scaffold. In this paper, we studied a bacteriophage selected through phage-display technol. The chosen bacteriophage acted as a building block for creating a carbon nanofiber-based electrode and as a new receptor/binding element that recognizes C-reactive protein (CRP) - one of the markers of inflammatory processes in the human body. The binding efficiency of the selected phage towards CRP is two orders of magnitude higher than in the wild type. We demonstrate that the phage-based sensor is selective against other proteins. Finally, we show that layer-by-layer methods are suitable for deposition of neg. charged phages (wild or CRP-binding) with pos. charged carbon nanofibers for electrode surface modification. A three-layered electrode was successfully used for mol. recognition of CRP, and the mol. interactions were studied using electrochem., biol., and optical methods, including microscopic and spectroscopic anal.
    35. 35
      Oloketuyi, S.; Bernedo, R.; Christmann, A.; Borkowska, J.; Cazzaniga, G.; Schuchmann, H. W.; Niedziółka-Jönsson, J.; Szot-Karpińska, K.; Kolmar, H.; de Marco, A. Native Llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection. Biosensors 2021, 11 (12), 496,  DOI: 10.3390/bios11120496
      There is no corresponding record for this reference.
    36. 36
      Beck, J. J.; Alenicheva, V.; Rahn, K. L.; Russo, M. J.; Baldo, T. A.; Henry, C. S. Evaluating the Performance of an Inexpensive, Commercially Available, NFC-Powered and Smartphone Controlled Potentiostat for Electrochemical Sensing. Electroanalysis 2023, 35 (6), e202200552  DOI: 10.1002/elan.202200552
      There is no corresponding record for this reference.
    37. 37
      Pungjunun, K.; Yakoh, A.; Chaiyo, S.; Siangproh, W.; Praphairaksit, N.; Chailapakul, O. Smartphone-Based Electrochemical Analysis Integrated with NFC System for the Voltammetric Detection of Heavy Metals Using a Screen-Printed Graphene Electrode. Microchim. Acta 2022, 189 (5), 191,  DOI: 10.1007/s00604-022-05281-x
      There is no corresponding record for this reference.
    38. 38
      Veggiani, G.; de Marco, A. Improved Quantitative and Qualitative Production of Single-Domain Intrabodies Mediated by the Co-Expression of Erv1p Sulfhydryl Oxidase. Protein Expression Purif. 2011, 79 (1), 111– 114,  DOI: 10.1016/j.pep.2011.03.005
      There is no corresponding record for this reference.
    39. 39
      Amatore, C.; Pebay, C.; Sella, C.; Thouin, L. Mass Transport at Microband Electrodes: Transient, Quasi-Steady-State, and Convective Regimes. ChemPhysChem 2012, 13 (6), 1562– 1568,  DOI: 10.1002/cphc.201100942
      There is no corresponding record for this reference.
    40. 40
      International, A. AOAC Peer-Verified Methods Program: Manual on Policies and Procedures; Association of Official Analytical Chemists, 1993.
      There is no corresponding record for this reference.
    41. 41
      Jönsson-Niedziolka, M.; Boonkaew, S. Electrochemical Data for NFC Smartphone-Based Electrochemical Microfluidic Device Integrated with Nanobody Recognition for C-Reactive Protein, RepOD – Repository for Open Data, 2024. https://doi.org/10.18150/M8JBP9 (accessed Feb 01, 2024).
      There is no corresponding record for this reference.

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    • Additional experimental details, scheme of the NFC potentiostat setup, cost breakdown of device, and comparison of literature CRP sensors (PDF)


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