PROTEOLYTIC FRAGMENTATION WITH HIGH SPECIFICITY OF MOUSE IMMUNOGLOBULIN-G - MAPPING OF PROTEOLYTIC CLEAVAGE SITES IN THE HINGE REGION

被引：65

作者：

YAMAGUCHI, Y ^{[1
]}

KIM, H ^{[1
]}

KATO, K ^{[1
]}

MASUDA, K ^{[1
]}

SHIMADA, I ^{[1
]}

ARATA, Y ^{[1
]}

机构：

[1] UNIV TOKYO,FAC PHARMACEUT SCI,BUNKYO KU,TOKYO 113,JAPAN

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1995年 / 181卷 / 02期

关键词：

IGG; MOUSE; PROTEASE; FRAGMENTATION; DIGESTION; SUBCLASS; HINGE REGION;

D O I：

10.1016/0022-1759(95)00010-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the results of fragmentation of mouse IgG by clostripain, lysyl endopeptidase, metalloendopeptidase, and V8 protease that have a narrower substrate specificity than papain and pepsin. A panel of mouse monoclonal switch variant antibodies with IgG1, IgG2a, and IgG2b subclasses were examined. Cleavage sites by these proteases were mapped on the hinge region of each of the IgG subclasses. It was demonstrated that lysyl endopeptidase can cleave the core hinge portion of IgG2a and IgG2b without perturbing the inter-chain disulfide bridges. Digestion products were successfully isolated by a combined use of protein A affinity chromatography and HPLC techniques. This is a first successful attempt of obtaining the F(ab')(2) fragment of IgG2b by proteolytic digestion.

引用

页码：259 / 267

页数：9

共 41 条

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