Journal
MICROCHIMICA ACTA
Volume 180, Issue 7-8, Pages 711-717Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-013-0974-y
Keywords
ELISA; Signal amplification; PolyHRP; Small molecule
Categories
Funding
- National Natural Science Foundation of China [30825027]
- German Ministry of Economics and Technology (via AiF)
- FEI (Forschungskreis der Ernahrungsindustrie e.V. Bonn)
- National Public Welfare Project for Agriculture [200903009]
- [AiF 381 ZN]
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We have developed a simple and sensitive competitive enzyme-linked immunosorbent assay (ELISA) to determine aflatoxin B1 (as a model small analyte) and using streptavidin-polymeric horseradish peroxidase complex (SApolyHRP) as a label for signal amplification. The performance of the assay was evaluated by comparing it with the classical indirect competitive ELISA using HRP labeled anti-mouse IgG as the tracer antibody. The results indicate that the SApolyHRP-based competitive ELISA exhibits a typically 2.4-fold steeper slope of the linear working range of the calibration curve compared to the monomeric HRP based classical ELISA, i.e., the sensitivity was increased. The SApolyHRP conjugate causes a typically 19-fold stronger signal generation in comparison to the traditional HRP labeled anti-mouse IgG at the same concentration (25 ng mL(-1)). Moreover, the SApolyHRP-based assay has a much wider linear range and a 3.8-fold better signal-to-noise ratio. Considering its simplicity, sensitivity and ease of operation, this competitive ELISA is considered to be a promising tool for small molecule immunodetection.
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