Aptasensor for quantification of leptin through PCR amplification of short DNA-aptamers

Cavallo, Francesca Romana, Mirza, Khalid B., de Mateo, Sara, Nikolic, Konstantin ORCID: https://orcid.org/0000-0002-6551-2977, Rodriguez-Manzano, Jesus and Toumazou, Christofer (2021) Aptasensor for quantification of leptin through PCR amplification of short DNA-aptamers. ACS Sensors, 6 (3). pp. 709-715.

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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.

Item Type: Article
Uncontrolled Keywords: aptamers, protein quantification, leptin, qPCR, target-induced disassociation
Subjects: Construction and engineering > Biomedical engineering
Related URLs:
Depositing User: Konstantin Nikolic
Date Deposited: 04 Mar 2021 10:08
Last Modified: 28 Aug 2021 07:14
URI: http://repository.uwl.ac.uk/id/eprint/7727

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