title: 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: Centre for Bioanalytical Sciences (CBAS), National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
authors: Chandra Kumar
authors: CK
authors: Dixit
authors: Sandeep Kumar
authors: SK
authors: Vashist
authors: Feidhlim T
authors: FT
authors: O'Neill
authors: Brian
authors: B
authors: O'Reilly
authors: Brian D
authors: BD
authors: MacCraith
authors: Richard
authors: R
authors: O'Kennedy
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D050197:Atherosclerosis / Q000175:diagnosis; D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008113:Liver Neoplasms / Q000175:diagnosis; D011108:Polymers / Q000737:chemistry; D012821:Silanes / Q000737:chemistry; D060749:alpha-2-HS-Glycoprotein
chemical_list: C556354:AHSG protein, human; D055910:Antibodies, Immobilized; D015415:Biomarkers; D001798:Blood Proteins; D011108:Polymers; D012821:Silanes; D060749:alpha-2-HS-Glycoprotein