Evaluation of Different Nonspecific Binding Blocking Agents Deposited Inside Poly(methyl methacrylate) Microfluidic Flow-CellsClick to copy article linkArticle link copied!
Abstract
Poly(methyl methacrylate) (PMMA) flow-cells containing microwells were deposited with different nonspecific binding blocking agents, namely, bovine serum albumin (BSA), cationic lipid (DOTAP:DOPE) and diethylene glycol dimethyl ether (DEGDME). Water contact angle (WCA) and atomic force microscope (AFM) measurements were carried out to confirm the successful depositions of BSA, DOTAP, and DEGDME onto the PMMA surfaces. Fluorescent intensity measurements were performed to evaluate the degree of nonspecific adsorption of Cy5-labeled anti-IgG proteins onto plain and oxygen plasma-treated (PT) PMMA flow-cells as well as PMMA flow-cells deposited with different above-mentioned blocking agents. We then employed a label-free detection method called total internal reflection ellipsometry (TIRE) to evaluate the stability of the deposited blocking agents inside the PMMA flow-cells. It was found that, while DOTAP:DOPE was the best agent for blocking the nonspecific adsorption, it could be removed from the PMMA surfaces of the flow-cells upon rinsing with phosphate buffered saline (PBS) and later deposited back onto the Au-coated glass sensing substrate of the TIRE. The removal of the blocking agents from PMMA surfaces and their deposition onto the sensing substrate were further manifested by measuring the kinetics and the amount of adsorbed anti-α-hCG proteins. Overall, the dry DEGDME coating by plasma-enhanced chemical vapor deposition (PECVD) showed very good blocking and excellent stability for subsequent assay inside the microwells. Our results could be useful when one considers what blocking agents should be used for PMMA-based microfluidic immunosensor or biosensor devices by looking at both the blocking efficiency and the stability of the blocking agent.
Cited By
This article is cited by 23 publications.
- Jan-Niklas Klatt, Tobias Hutzenlaub, Thomas Subkowski, Tanja Müller, Stefan Hennig, Roland Zengerle, Nils Paust. Blocking Protein Adsorption in Microfluidic Chips by a Hydrophobin Coating. ACS Applied Polymer Materials 2021, 3
(7)
, 3278-3286. https://doi.org/10.1021/acsapm.0c01301
- Filip Kunc, Colin J. Moore, Rachel E. Sully, Andrew J. Hall, Vladimir Gubala. Polycarboxylated Dextran as a Multivalent Linker: Synthesis and Target Recognition of the Antibody–Nanoparticle Bioconjugates in PBS and Serum. Langmuir 2019, 35
(14)
, 4909-4917. https://doi.org/10.1021/acs.langmuir.8b03833
- Yuqing Lin, Kangyu Liu, Chao Wang, Linbo Li, and Yuxin Liu . Electrochemical Immunosensor for Detection of Epidermal Growth Factor Reaching Lower Detection Limit: Toward Oxidized Glutathione as a More Efficient Blocking Reagent for the Antibody Functionalized Silver Nanoparticles and Antigen Interaction. Analytical Chemistry 2015, 87
(16)
, 8047-8051. https://doi.org/10.1021/acs.analchem.5b01834
- Karen Scida, Bingling Li, Andrew D. Ellington, and Richard M. Crooks . DNA Detection Using Origami Paper Analytical Devices. Analytical Chemistry 2013, 85
(20)
, 9713-9720. https://doi.org/10.1021/ac402118a
- King Hang Aaron Lau, Chunlai Ren, Tadas S. Sileika, Sung Hyun Park, Igal Szleifer, and Phillip B. Messersmith . Surface-Grafted Polysarcosine as a Peptoid Antifouling Polymer Brush. Langmuir 2012, 28
(46)
, 16099-16107. https://doi.org/10.1021/la302131n
- Vladimir Gubala, Leanne F. Harris, Antonio J. Ricco, Ming X. Tan, and David E. Williams . Point of Care Diagnostics: Status and Future. Analytical Chemistry 2012, 84
(2)
, 487-515. https://doi.org/10.1021/ac2030199
- Mustafa Oguzhan Caglayan. Ellipsometric biosensors. 2023, 197-237. https://doi.org/10.1016/B978-0-323-88431-0.00016-8
- Yali Chen, Ling Xia, Xiaohua Xiao, Gongke Li. Enhanced capillary zone electrophoresis in cyclic olefin copolymer microchannels using the combination of dynamic and static coatings for rapid analysis of carnosine and niacinamide in cosmetics. Journal of Separation Science 2022, 45
(12)
, 2045-2054. https://doi.org/10.1002/jssc.202101007
- Borys Snopok, Arwa Laroussi, Clodomiro Cafolla, Kislon Voïtchovsky, Tetyana Snopok, Vladimir M. Mirsky. Gold surface cleaning by etching polishing: Optimization of polycrystalline film topography and surface functionality for biosensing. Surfaces and Interfaces 2021, 22 , 100818. https://doi.org/10.1016/j.surfin.2020.100818
- Tian Yang, Shengfu Chen, Xiaolong He, Huaizu Guo, Xiaqing Sun. How to convincingly measure low concentration samples with optical label-free biosensors. Sensors and Actuators B: Chemical 2020, 306 , 127568. https://doi.org/10.1016/j.snb.2019.127568
- Ruwen Tan, Jin Yoo, Yeongseon Jang. Engineering Approaches to Create Antibacterial Surfaces on Biomedical Implants and Devices. 2020, 313-340. https://doi.org/10.1007/978-3-030-34475-7_14
- Richard O’Kennedy, Jenny Fitzgerald, Arabelle Cassedy, Aoife Crawley, Xin Zhang, Sandro Carrera. Applications of antibodies in microfluidics-based analytical systems: challenges and strategies for success. Journal of Micromechanics and Microengineering 2018, 28
(6)
, 063001. https://doi.org/10.1088/1361-6439/aab225
- Fupeng Liang, Yi Qiao, Mengqin Duan, An Ju, Na Lu, Junji Li, Jing Tu, Zuhong Lu. Fabrication of a microfluidic chip based on the pure polypropylene material. RSC Advances 2018, 8
(16)
, 8732-8738. https://doi.org/10.1039/C7RA13334K
- Ayesha Kausar. Adhesion, morphology, and heat resistance properties of polyurethane coated poly(methyl methacrylate)/fullerene-C
60
composite films. Composite Interfaces 2017, 24
(7)
, 649-662. https://doi.org/10.1080/09276440.2017.1257251
- Xin Zhang, Shou-Nian Ding. Graphite paper-based bipolar electrode electrochemiluminescence sensing platform. Biosensors and Bioelectronics 2017, 94 , 47-55. https://doi.org/10.1016/j.bios.2017.02.033
- Isabel Álvarez-Martos, Rebeca Alonso-Bartolomé, Verónica Mulas Hernández, Adela Anillo, Agustín Costa-García, Francisco Javier García Alonso, María Teresa Fernández-Abedul. Poly(glycidyl methacrylate) as a tunable platform of modifiers for microfluidic devices. Reactive and Functional Polymers 2016, 100 , 89-96. https://doi.org/10.1016/j.reactfunctpolym.2016.01.002
- I. Almeida, J.T. Marquês, W. Liu, Y. Niu, R.F.M. de Almeida, G. Jin, A.S. Viana. Phospholipid/cholesterol/decanethiol mixtures for direct assembly of immunosensing interfaces. Colloids and Surfaces B: Biointerfaces 2015, 136 , 997-1003. https://doi.org/10.1016/j.colsurfb.2015.10.048
- Rashid Zakeri, Joseph R. Basore, Lane A. Baker. Modulated fluorescence detection with microelectromagnetic traps. Analytical Methods 2015, 7
(6)
, 2273-2277. https://doi.org/10.1039/C4AY02828G
- David Porciani, Giovanni Signore, Laura Marchetti, Paolo Mereghetti, Riccardo Nifosì, Fabio Beltram. Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor. Molecular Therapy - Nucleic Acids 2014, 3 , e144. https://doi.org/10.1038/mtna.2013.71
- Namsoo Kim, Yong-Jin Cho. Optimization of Fluoroimmunoassay against C-Reactive Protein Exploiting Immobilized-antigen Glass Slide. Journal of Fluorescence 2013, 23
(2)
, 243-249. https://doi.org/10.1007/s10895-012-1140-3
- Vladimir Gubala, Jonathan Siegrist, Ruairi Monaghan, Brian O’Reilly, Ram Prasad Gandhiraman, Stephen Daniels, David E. Williams, Jens Ducrée. Simple approach to study biomolecule adsorption in polymeric microfluidic channels. Analytica Chimica Acta 2013, 760 , 75-82. https://doi.org/10.1016/j.aca.2012.11.030
- Nam Cao Hoai Le, Vladimir Gubala, Eoin Clancy, Thomas Barry, Terry J Smith, David E. Williams. Ultrathin and smooth poly(methyl methacrylate) (PMMA) films for label-free biomolecule detection with total internal reflection ellipsometry (TIRE). Biosensors and Bioelectronics 2012, 36
(1)
, 250-256. https://doi.org/10.1016/j.bios.2012.04.032
- Gowri Manickam, Ram Gandhiraman, Rajani K. Vijayaraghavan, Laura Kerr, Colin Doyle, David E. Williams, Stephen Daniels. Protection and functionalisation of silver as an optical sensing platform for highly sensitive SPR based analysis. The Analyst 2012, 137
(22)
, 5265. https://doi.org/10.1039/c2an35826c
Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.
Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.
The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.