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Aptasensor for Quantification of Leptin Through PCR Amplification of Short DNA-Aptamers

  • Francesca Romana Cavallo*
    Francesca Romana Cavallo
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
    *Email: [email protected]
  • Khalid B. Mirza*
    Khalid B. Mirza
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
    Department of Biotechnology and Medical Engineering, National Institute of Technology, Rourkela, Odisha 769008, India
    *Email: [email protected]
  • Sara de Mateo
    Sara de Mateo
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
  • Konstantin Nikolic
    Konstantin Nikolic
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
    School of Computing and Engineering, University of West London, London W5 5RF, United Kingdom
  • Jesus Rodriguez-Manzano
    Jesus Rodriguez-Manzano
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
    Department of Infectious Disease, Imperial College London, London SW7 2AZ, United Kingdom
  • , and 
  • Christofer Toumazou*
    Christofer Toumazou
    Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United Kingdom
    *Email: [email protected]
Cite this: ACS Sens. 2021, 6, 3, 709–715
Publication Date (Web):March 2, 2021
https://doi.org/10.1021/acssensors.0c02605
Copyright © 2021 American Chemical Society
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Supporting Info (1)»

Abstract

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Protein quantification is traditionally performed through enzyme-linked immunosorbent assay (ELISA), which involves long preparation times. To overcome this, new approaches use aptamers as an alternative to antibodies. In this paper, we present a new approach to quantify proteins with short DNA aptamers through polymerase chain reaction (PCR) resulting in shorter protocol times with comparatively improved limits of detection. The proposed method includes a novel way to quantify both the target protein and the corresponding short DNA-aptamers simultaneously, which also allows us to fully characterize the performance of aptasensors. Human leptin is used as a target protein to validate this technique, because it is considered an important biomarker for obesity-related studies. In our experiments, we achieved the lowest limit of detection of 100 pg/mL within less than 2 h, a limit affected by the dissociation constant of the leptin aptamer, which could be improved by selecting a more specific aptamer. Because of the simple and inexpensive approach, this technique can be employed for Lab-On-Chip implementations and for rapid “on-site” quantification of proteins.

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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acssensors.0c02605.

  • Additional results for proposed aptasensor and description of the aptasensor with beads (PDF)

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Cited By

This article is cited by 6 publications.

  1. Fangfang Xia, Axin He, Haitao Zhao, Yang Sun, Qiao Duan, Sk Jahir Abbas, Jianjun Liu, Zeyu Xiao, Weihong Tan. Molecular Engineering of Aptamer Self-Assemblies Increases in Vivo Stability and Targeted Recognition. ACS Nano 2022, 16 (1) , 169-179. https://doi.org/10.1021/acsnano.1c05265
  2. Zeinab Rouhbakhsh, Tsung Yang Ho, Asma Verdian, Chih-Hsin Chen. Detection of okadaic acid using a liquid crystal-based aptasensor by exploiting the signal enhancement effect of gold nanoparticles. Biosensors and Bioelectronics: X 2022, 11 , 100148. https://doi.org/10.1016/j.biosx.2022.100148
  3. Francesca Romana Cavallo, Khalid Baig Mirza, Sara de Mateo, Luca Miglietta, Jesus Rodriguez-Manzano , Konstantin Nikolic, Christofer Toumazou. A Point-of-Care Device for Fully Automated, Fast and Sensitive Protein Quantification via qPCR. Biosensors 2022, 12 (7) , 537. https://doi.org/10.3390/bios12070537
  4. Cem ERKMEN, Gözde AYDOĞDU TIĞ, Bengi USLU. First label-free impedimetric aptasensor based on Au NPs/TiO2 NPs for the determination of leptin. Sensors and Actuators B: Chemical 2022, 358 , 131420. https://doi.org/10.1016/j.snb.2022.131420
  5. Yuan Fan, Shaobo Ou‐yang, Dong Zhou, Junchao Wei, Lan Liao. Biological applications of chiral inorganic nanomaterials. Chirality 2022, 34 (5) , 760-781. https://doi.org/10.1002/chir.23428
  6. İnci Uludağ, Mustafa Kemal Sezgintürk. A direct and simple immobilization route for immunosensors by CNBr activation for covalent attachment of anti-leptin: obesity diagnosis point of view. 3 Biotech 2022, 12 (1) https://doi.org/10.1007/s13205-021-03096-w

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