固定化木瓜蛋白酶水解HCG抗体及其片段的纯化  

Hydrolysis of HCG Monoclonal Antibodies with Immobilized Papain and Purifi cation of Its Fragments

作　　者：易喻[1] 高虹[1] 陈建澍[1] 朱克寅 应国清[1] 

机构地区：[1]浙江工业大学药学院,浙江杭州310014 [2]杭州博林生物技术有限公司,浙江杭州310018

出　　处：《中国医药工业杂志》2014年第9期826-829,共4页Chinese Journal of Pharmaceuticals

基　　金：浙江省科技计划项目(2012C23076)

摘　　要：采用温敏载体NHS-PNIPAM固定化木瓜蛋白酶,水解β-HCG单克隆抗体制备Fab片段,并优化水解工艺。结果表明:30℃,在pH 7.0的磷酸盐缓冲液中(含10 mmol/L半胱氨酸,2 mmol/L EDTA),加酶量40 u·(mg抗体)-1,转速180 r/min,反应3 h,β-HCG抗体的转化率大于98.0%,主要生成目的产物Fab片段。酶解产物通过Protein L亲和色谱法分离纯化,得到的Fab片段回收率为74.5%,纯度为98.6%。HCG monoclonal antibody was hydrolyzed by papain immobilized in NHS-PNIPAM and the hydrolysis process was optimized to prepare Fab fragments. The results showed that 1 mg HCG antibodies were hydrolyzed to Fab fragments with conversion rate over 98.0 % by 40 u immobilized papain active for 3 h at 30℃, pH 7.0, 180 r/min in 1 ml phosphate buffer solution （pH 7.0） containing 10 mmol/L cysteine and 2 mmol/L EDTA. Fab fragments were purified by Protein L affinity chromatography from the hydrolysis mixture. The purity of the obtained Fab fragments was 98.6 % with the recovery of 74.5 %.

关 键 词：HCG抗体 水解 木瓜蛋白酶 纯化 FAB片段 

分 类 号：Q511[生物学—生物化学]