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  • Title: Production of a recombinant Fab in Pichia pastoris from a Monocistronic expression vector.
    Author: Burtet RT, Santos-Silva MA, Buss GA, Moraes LM, Maranhão AQ, Brigido MM.
    Journal: J Biochem; 2007 Dec; 142(6):665-9. PubMed ID: 18037691.
    Abstract:
    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.
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