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Determination of the platelet-derived growth factor BB by a sandwich format thrombin-linked aptamer assay on a microplate

Author:
Guo, Limin
Source:
Analytical methods 2019 v.11 no.28 pp. 3532-3537
ISSN:
1759-9679
Subject:
DNA probes, analytical methods, antibodies, catalytic activity, detection limit, oligonucleotides, platelet-derived growth factor, rapid methods, thrombin
Abstract:
Microplate based assays have advantages such as rapid detection and high throughput analysis. Here, we reported a thrombin linked aptamer assay (TLAA) on a microplate in sandwich format for the detection of platelet-derived growth factor BB (PDGF-BB) using thrombin as the enzyme label. In our assay, the target protein PDGF-BB was captured by anti-PDGF-BB antibody coated on the microplate and sandwiched with a DNA probe containing an anti-PDGF-BB aptamer sequence and anti-thrombin aptamer sequence. Thrombin was labeled on the sandwich complex through aptamer–thrombin interaction. Finally, the thrombin catalyzed the conversion of the substrate to a detectable product, generating signals for final PDGF-BB detection. The sandwich format of the TLAA on a microplate allowed the selective detection of PDGF-BB with a detection limit of 0.25 nM.
Agid:
6539526