Production of a recombinant fab in Pichia pastoris from a monocistronic expression vector

被引：8

作者：

Burtet, Rafael Trindade ^{[1
,2
]}

Santos-Silva, Marcos Antonio ^{[1
]}

Antonio, Guilherme ^{[1
]}

Buss, Marques ^{[1
]}

Pepe Moraes, Lidia Maria ^{[1
]}

Maranhao, Andrea Queiroz ^{[1
,2
]}

Brigido, Marcelo Macedo ^{[1
,2
]}

机构：

[1] Univ Brasilia, BR-70910900 Brasilia, DF, Brazil

[2] Millennium Inst, Inst Invest Immunol, MCT, Brasilia, DF, Brazil

来源：

JOURNAL OF BIOCHEMISTRY | 2007年 / 142卷 / 06期

关键词：

antibody engineering; fab expression; kex; Pichia pastoris; SINGLE-CHAIN FV; ANTIBODY FRAGMENTS; ESCHERICHIA-COLI; Z-DNA; SECRETION; TOXIN; GENES;

D O I：

10.1093/jb/mvm226

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Recombinant Fab is usually expressed using dicistronic vectors producing the heavy and light chains separately. We developed an improved vector for Fab fragment expression in Pichia pastoris, which allows a stoichiometric expression of both chains based on a monocistronic arrangement. The protein is produced as a unique polypeptide harbouring a KEX2 processing site between both chains. After KEX cleavage, a correctly folded mature Fab is formed. The produced recombinant protein is characterized as a heterodimeric functional Fab. The vector described is a new tool for the proper expression of antibody fragments or any heterodimeric polypeptides.

引用

页码：665 / 669

页数：5

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