Development of a high sensitivity rapid sandwich ELISA procedure and its comparison with the conventional approach.

Abstract

A highly sensitive and rapid sandwich enzyme-linked immunosorbent assay (ELISA) procedure was developed for the detection of human fetuin A/AHSG (alpha2-HS-glycoprotein), a specific biomarker for hepatocellular carcinoma and atherosclerosis. Anti-human fetuin A antibody was immobilized on aminopropyltriethoxysilane-mediated amine-functionalized microtiter plates using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride and N-hydroxysulfosuccinimide-based heterobifunctional cross-linking. The analytical sensitivity of the developed assay was 39 pg/mL, compared to 625 pg/mL for the conventional assay. The generic nature of the developed procedure was demonstrated by performing human fetuin A assays on different polymeric matrixes, i.e., polystyrene, poly(methyl methacrylate), and polycyclo-olefin (Zeonex), in a modified microtiter plate format. Thus, the newly developed procedure has considerable advantages over the existing method.

• Authors: Chandra Kumar Dixit, Sandeep Kumar Vashist, Feidhlim T O'Neill, Brian O'Reilly, Brian D MacCraith, Richard O'Kennedy
• Journal: Analytical chemistry (Anal Chem) Vol. 82 Issue 16 Pg. 7049-52 (Aug 15 2010) ISSN: 1520-6882 [Electronic] United States
• PMID: 20704394 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• AHSG protein, human
• Antibodies, Immobilized
• Biomarkers
• Blood Proteins
• Polymers
• Silanes
• alpha-2-HS-Glycoprotein

Topics

• Antibodies, Immobilized (chemistry, immunology)
• Atherosclerosis (diagnosis)
• Biomarkers (analysis)
• Blood Proteins (analysis)
• Carcinoma, Hepatocellular (diagnosis)
• Enzyme-Linked Immunosorbent Assay (methods)
• Humans
• Liver Neoplasms (diagnosis)
• Polymers (chemistry)
• Silanes (chemistry)
• alpha-2-HS-Glycoprotein