Preparation of egg yolk antibodies against human isomaltase and determination of their biological activities / 医学研究生学报

ABSTRACT

Objective Antibody drugs are one of the hot topics in biomedical research.This study aims to develop egg yolk antibodies (IgYs) against human isomaltase and determine their biological activities.Methods The purified recombinant isomaltase protein was used as an antigen to immunize egg-laying hens in combination with complete Freund adjuvant (CFA).Anti-isomahase IgYs were extracted by water dilution-sodium sulfate extraction assay and further analyzed for their purity,specificity,titer and stability by SDS-PAGE,Western blot and ELISA respectively,and their inhibitory effect on human alpha-glycosidase enzymes was evaluated by the PNPG method.Results Anti-isomaltase IgYs were obtained,with a titer of 1 ∶ 12800,capable of specifically binding human isomaltase,and with a good thermal stability,acid/alkali stability and pepsin resistance.Conclusion Anti-human isomaltase IgYs were successfully prepared,which may provide an experimental ground for further investigation of oral antihyperglycemic agents for type Ⅱ diabetes mellitus.