Production of a recombinant fab in Pichia pastoris from a monocistronic expression vector
Production of a recombinant fab in Pichia pastoris from a monocistronic expression vector
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DOI：
10.1093/jb/mvm226
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发表时间：
2007-12-01
影响因子：
2.7
通讯作者：
Brigido, Marcelo Macedo
中科院分区：
文献类型：
--
作者：
Burtet, Rafael Trindade;Santos-Silva, Marcos Antonio;Brigido, Marcelo Macedo
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.
