Aptasensor for Quantification of Leptin Through PCR Amplification of Short DNA-Aptamers
- Francesca Romana Cavallo*Francesca Romana Cavallo*Email: [email protected]Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomMore by Francesca Romana Cavallo,
- Khalid B. Mirza*Khalid B. Mirza*Email: [email protected]Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomDepartment of Biotechnology and Medical Engineering, National Institute of Technology, Rourkela, Odisha 769008, IndiaMore by Khalid B. Mirza,
- Sara de MateoSara de MateoCentre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomMore by Sara de Mateo,
- Konstantin NikolicKonstantin NikolicCentre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomSchool of Computing and Engineering, University of West London, London W5 5RF, United KingdomMore by Konstantin Nikolic,
- Jesus Rodriguez-ManzanoJesus Rodriguez-ManzanoCentre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomDepartment of Infectious Disease, Imperial College London, London SW7 2AZ, United KingdomMore by Jesus Rodriguez-Manzano, and
- Christofer Toumazou*Christofer Toumazou*Email: [email protected]Centre for Bio-Inspired Technology, Imperial College London, London SW7 2AZ, United KingdomMore by Christofer Toumazou
Abstract
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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