Development of a high sensitivity rapid sandwich ELISA procedure and its comparison with the conventional approach.
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.
Dixit
Chandra Kumar
CK
Centre for Bioanalytical Sciences (CBAS), National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
Vashist
Sandeep Kumar
SK
O'Neill
Feidhlim T
FT
O'Reilly
Brian
B
MacCraith
Brian D
BD
O'Kennedy
Richard
R
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
C556354:AHSG protein, human; D055910:Antibodies, Immobilized; D015415:Biomarkers; D001798:Blood Proteins; D011108:Polymers; D012821:Silanes; D060749:alpha-2-HS-Glycoprotein