Abstract:In this study,dextran neoglycoprotein antigens were prepared evaluated and finally obtained dextran polyclonal antibodies,and an enzyme-linked immunosorbent assay(Elisa) detection method was developed according to Elisa theory. The method was put to use for quantitative analysis of dextran in practical samples. 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 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. The best preparation conditions were obtained(ndextran /noxidant of NaIO4 =1/120,the reaction time of 24 h),and the antigen with best combination with standard was selected. 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 indirect competitive Elisa method was put to use for quantitative analysis of dextran T40 in commercial sugar. The different preparation condition of dextran neoglycoprotein antigen has effect on its immunocompetence. The developed Elisa detection method with good linear and accuracy was practical for dextran determination.
