A Highly-Sensitive Colorimetric Assay Method for Antibody Array Based on an Tyramide Signal Amplification System

摘要

In this work, a highly-sensitive and cost-effective detection approach based on the integration of tyramide signal amplification with a silver enhancement method (SEM-TSA) has been developed successfully. To demonstrate the feasibility of this approach, human IgG as a model target protein was employed and its concentration was assayed based on colorimetric detection. The analytical parameters including the concentrations of detection antibody, streptavidin-horseradish peroxidase, biotinyl tyramide, and streptavidin-nanogold were systematically optimized. The quantitative analysis was performed and a dynamic range was obtained from 0.18Â ng/mL to 39.1Â ng/mL, while no detectable images could be observed when the silver enhancement method (SEM) without TSA was used. The detection limits were 0.18Â ng/mL and 21Â ng/mL for SEM-TSA and SEM, respectively. The results showed that sensitivity of the presented colorimetric assay significantly increased by two-orders of magnitude. In addition, this method has been successfully applied in analyzing normal human serum samples. The results suggested the colorimetric detection method based on TSA-SEM has promising potential applications in biomedical analysis and clinical diagnosis.View full textDownload full textKeywordsAntibody arrays, Colorimetric assay, Silver enhancement method, Tyramide signal amplificationRelated var addthis_config = { ui_cobrand: "Taylor & Francis Online", services_compact: "citeulike,netvibes,twitter,technorati,delicious,linkedin,facebook,stumbleupon,digg,google,more", pubid: "ra-4dff56cd6bb1830b" }; Add to shortlist Link Permalink http://dx.doi.org/10.1080/00032719.2011.633183