Sandeep Kumar Vashist - Development of a high sensitivity rapid sandwich ELISA procedure and its comparison with the conventional approach

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    ANALYTICAL CHEMISTRY,   82(16),   7049-52, 2010.


  A highly sensitive and rapid sandwich enzyme-linked immunosorbent   assay (ELISA) procedure was developed for the detection of human   fetuin A/AHSG (α2-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.

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