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Chemiluminescent and Colorimetric Aptamer-Based Assays of Human α-Thrombin.
- Source :
-
Analytical Letters . 2020, Vol. 53 Issue 1, p140-151. 12p. 1 Diagram, 5 Graphs. - Publication Year :
- 2020
-
Abstract
- A microplate-based sandwich assay for the determination of α-human thrombin (HTb) was developed. Fluorescein-modified 29-mer thrombin binding aptamer (FAM-TBA29) and biotinylated 15-mer thrombin binding aptamer (Bio-TBA15) reacting with different exosites of HTb were used as the biorecognition components in the assay. FAM-TBA29 (capture aptamer) was immobilized using its interaction with anti-fluorescein antibody adsorbed on the microplate surface. As a sensitive signaling system, a combination of Bio-TBA15 and streptavidin-polyHRP conjugate was used. Under the optimized conditions, the detection limit for HTb was 1.4 nM; this value was the same in both the colorimetric and the chemiluminescent assays. The replacement of colorimetry for HRP measurement with chemiluminescence increased the assay sensitivity from 0.06 to 1.7 × 106 nM−1 that clearly demonstrated advantage of the latter approach. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00032719
- Volume :
- 53
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Analytical Letters
- Publication Type :
- Academic Journal
- Accession number :
- 139617834
- Full Text :
- https://doi.org/10.1080/00032719.2019.1640718