Thrombin-linked aptamer assay for detection of platelet derived growth factor BB on magnetic beads in a sandwich format

TALANTA

卷:158

Article

Guo, Limin^1,3  Zhao, Qiang^1,2 

[1] Shanxi Univ, Inst Environm Sci, Coll Chem & Chem Engn, Taiyuan 030006, Peoples R China

[2] Chinese Acad Sci, Res Ctr Ecoenvironm Sci, State Key Lab Environm Chem & Ecotoxicol, Beijing 100085, Peoples R China

[3] Shanxi Univ Tradit Chinese Med, Sch Chinese Mat Med, Taiyuan 030024, Peoples R China

关键词: Aptamer;    Thrombin;    Enzyme label;    Protein detection;    Affinity binding;    Platelet derived growth factor BB;   

DOI  :  10.1016/j.talanta.2016.05.037

来源: Elsevier

【 摘 要 】

Here we describe a thrombin-linked aptamer assay (TLAA) for protein by using thrombin as an enzyme label, harnessing enzyme activity of thrombin and aptamer affinity binding. TLAA converts detection of specific target proteins to the detection of thrombin by using a DNA sequence that consists of two aptamers with the first aptamer binding to the specific target protein and the second aptamer binding to thrombin. Through the affinity binding, the thrombin enzyme is labeled on the protein target, and thrombin catalyzes the hydrolysis of small peptide substrate into product, generating signals for quantification. As a proof of principle, we show a sandwich TLAA for platelet derived growth factor BB (PDGF-BB) by using anti-PDGF-BB antibody coated on magnetic beads and an oligonucleotide containing the aptamer for PDGF-BB and the aptamer for thrombin. The binding of PDGF-BB to both the antibody and the aptamer results in labeling the complex with thrombin. We achieved detection of PDGF-BB at 16 pM. This TLAA contributes a new application of thrombin and its aptamer in bioanalysis, and shows potentials in assay developments. (C) 2016 Elsevier B.V. All rights reserved.

【 授权许可】

Free