title: Preparation and Characteristic of Dextran-BSA Antibody and Establishment of its ELISA Immunoassay.
abstract: The enzyme-linked immunosorbent assay (ELISA) is a potential tool for the determination of dextran. In this study, dextran neoglycoprotein antigens were prepared by Reductive Amination method, and were confirmed by SDS-PAGE and free amino detection. The impact factors such as different oxidation degree of dextran, the conjugate reaction time to BSA were investigated. The best preparation conditions were obtained (n(dextran)/n(oxidant) of NaIO4 = 1/120, the reaction time of 24 h), and the antigen with best combination with standard was obtained. The antigens interacted with standard antibody and were evaluated through ELISA. The immunogen was immunized with white rabbits to obtained antibody, respectively. A general and broad class-specific ELISA immunoassay was developed for dextran detection according to ELISA theory. The optimized conditions of assay used coating antigen at 10 μg/mL, reaction time of antibody and rabbit-anti-bovine IgG in 45 min, blocking reagents with 5% calf serum. The developed ELISA detection method with good linear and accuracy was put to use for quantitative analysis of dextran T40 in commercial sugarpractical for detection of dextran.
authors.affiliation: a Faculty of Light and Chemical Engineering , Guangdong University of Technology , Guangzhou , China.
authors.forename: Zhen-ming
authors.initials: ZM
authors.lastname: Xie
authors.forename: Lin
authors.initials: L
authors.lastname: Yu
authors.forename: Li-sha
authors.initials: LS
authors.lastname: Fang
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D002417:Cattle; D003911:Dextrans / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology*
chemical_list: D000941:Antigens; D003911:Dextrans; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine
keywords: ELISA; dextran; neoglycoprotein antigen; polyclonal antibody; preparation