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小鼠抗天花粉蛋白IgE型单抗的酶解及其Fab的制备
引用本文:何贤辉,柯一保,孙汛,蔡迎春,季永镛,聂慧玲.小鼠抗天花粉蛋白IgE型单抗的酶解及其Fab的制备[J].中国生物化学与分子生物学报,1995,11(6):697-701.
作者姓名:何贤辉  柯一保  孙汛  蔡迎春  季永镛  聂慧玲
作者单位:中国科学院上海细胞生物学研究所
基金项目:“863-103”国家高技术资金资助
摘    要: 研究了Papain及Trypsin裂解小鼠抗天花粉蛋白IgE单抗的条件及Fab的制备。Papain和Trypsin两者都可产生F(ab′)_2,分子量在150~160kD左右;经Papain裂解的主要产物中还有Fab,分子量72kD,可通过凝胶过滤获得纯的Fab。而Trypsin裂解物经DTT还原、碘乙酰胺烷化虽然也可得到Fab′(t),但不易纯化;可见,要制备Fab以采用Papain裂解为好,而制备F(ab′)_2则可采用Trypsin裂解。这二个酶的裂解速度是Trypsin大于Papain。

关 键 词:IgE  蛋白酶解  木瓜蛋白酶  胰蛋白酶  Fab
收稿时间:1995-12-20

Proteolysis of Mouse Anti-TCS IgE Monoclonal Antibody and Preparation of its Fab
He,Xian-Hui,Ke,Yi-Bao,Sun,Xun,Cai,Ying-Chun Ji,Yong-Yong,Nie,Hui-Ling.Proteolysis of Mouse Anti-TCS IgE Monoclonal Antibody and Preparation of its Fab[J].Chinese Journal of Biochemistry and Molecular Biology,1995,11(6):697-701.
Authors:He  Xian-Hui  Ke  Yi-Bao  Sun  Xun  Cai  Ying-Chun Ji  Yong-Yong  Nie  Hui-Ling
Institution:(Shanghai Institute of Cell Biology,Academia Sinica,Shanghai 200031
Abstract:The proteolysis of mouse IgE with papain and trypsin were investigated. F(ab')_2 could be produced by these two enzymes and its molecular weighgt was about 150-160 kD;Fab was also presented in the papain digestion as a main product with the molecular weight of 72 kD,which may be purified by gel filtration. Fab' could be obtained from the trypsin digestion after being reduced with DTT and alkylated with iodoacetamide,but it was difficult to get relatively pure Fab'(t). The molecular weights of heavy chain fragments in F(ab')_2 and Fab were 53. 5 kD and 37. 5 kD respectively. Both of them also contained intact light chains(27 kD). In conclusion, papain is the enzyme of choice for preparing Fab,while trypsin may be used for the preparation of F(ab')_2.Rate of digestion of papain was slower than that of trypsin.
Keywords:IgE  Proteolytic digestion  Papain  Trypsin  Fab
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