抗人异麦芽糖酶卵黄抗体的研制及活性测定  

Preparation of egg yolk antibodies against human isomaltase and determination of their biological activities

作　　者：王新颖[1] 王丹[1] 冯昆[1] 葛正龙[2] 周鹤峰[1] 邵敏[1] 

机构地区：[1]遵义医学院珠海校区生物工程系,珠海理学硕士研究生519041 [2]遵义医学院生物化学教研室,遵义563003

出　　处：《医学研究生学报》2018年第2期137-141,共5页Journal of Medical Postgraduates

基　　金：贵州省科技计划课题[黔科合SY字(2012)3091]

摘　　要：目的抗体药物是目前生物医药的研究热点问题之一。文章旨在研制抗人异麦芽糖酶(hISO)卵黄抗体(IIgY)并检测其生物学活性。方法将hISO重组蛋白作为抗原与弗式佐剂充分乳化后免疫蛋鸡,水稀释-硫酸钠提取法提取抗人I-IgY,分别通过SDS-PAGE、Western blot和ELISA等实验技术分析该抗体的纯度、特异性、效价和稳定性,PNPG法测定该抗体对α-葡萄糖苷酶的体外抑制效果。结果获得了抗人I-IgY,其效价最高可以达1∶12 800,该抗体可特异性结合hISO重组蛋白,且具有较好的热稳定性、酸碱稳定性及胃蛋白酶抗性。结论成功制备了抗人I-IgY,为后续对2型糖尿病口服降糖药物的研制奠定实验基础。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-isomaltase 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-isomahase IgYs were obtained, with a titer of 1 ： 12 800, capable of specifically binding human iso- maltase, 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.

关 键 词：异麦芽糖酶 卵黄抗体 效价 稳定性 特异性 

分 类 号：R392.11[医药卫生—免疫学]