SYNTHESIS AND EXPRESSION IN ESCHERICHIA-COLI OF CISTRONIC DNA ENCODING AN ANTIBODY FRAGMENT SPECIFIC FOR A SALMONELLA SEROTYPE-B O-ANTIGEN

被引：24

作者：

ANAND, NN ^{[1
]}

DUBUC, G ^{[1
]}

PHIPPS, J ^{[1
]}

MACKENZIE, CR ^{[1
]}

SADOWSKA, J ^{[1
]}

YOUNG, NM ^{[1
]}

BUNDLE, DR ^{[1
]}

NARANG, SA ^{[1
]}

机构：

[1] NATL RES COUNCIL CANADA,INST BIOL SCI,OTTAWA K1A 0R6,ONTARIO,CANADA

来源：

GENE | 1991年 / 100卷

关键词：

RECOMBINANT DNA; SYNTHON; FAB-GENE ASSEMBLY; POLYSACCHARIDE ANTIGEN;

D O I：

10.1016/0378-1119(91)90347-E

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A 1460-bp DNA encoding the two chains of the antigen-binding fragment (Fab) portion of a monoclonal antibody have been chemically synthesized and expressed in Escherichia coli. The antibody, Se155-4, is specific for a Salmonella serogroup B O-antigen and its crystal structure is under investigation. The genes were synthesized according to a strategy that allows for easy manipulation in genetic engineering studies of the Fab-binding site. Each gene is preceded by the ompA secretory signal and a ribosome-binding site, and has been expressed from the two-cistron DNA under the control of the lac promoter. Active Fab of 50 kDa with an inter-chain disulfide bond has been isolated from the periplasm of E. coli in a one-step affinity purification in high yield (2-mu-g/ml of cells). The bacterially produced Fab is a active as purified mouse Fab in antigen-binding and competitive immunoassays. This is the first example of a completely synthetic Fab gene and provides an ideal system to probe the nature of antigen binding by anti-carbohydrate antibodies.

引用

页码：39 / 44

页数：6

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