Skip Navigation



Journal of Biochemistry Advance Access published online on November 24, 2007
Journal of Biochemistry, doi:10.1093/jb/mvm226
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
142/6/665    most recent
mvm226v2
mvm226v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Burtet, R. T.
Right arrow Articles by Brigido, M. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Burtet, R. T.
Right arrow Articles by Brigido, M. M.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Japanese Biochemical Society

Rapid Communication

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

Rafael Trindade Burtet1,2, Marcos Antônio Santos-Silva1, Guilherme Antônio Marques Buss1, Lidia Maria Pepe Moraes1, Andrea Queiroz Maranhão1,2 and Marcelo Macedo Brigido1,2,*

1Universidade de Brasília, 70910-900, Brasília, DF, Brazil
2Instituto de Investigação em Imunologia

*Corresponding author: Dr. Marcelo Macedo Brigido, Departamento de Biologia Celular-IB, Universidade de Brasília, 70.910-900 Brasília, DF, Brasil,, Tel. 55 (61) 3072423; Fax. 55 (61) 3498411; E-mail: brigido{at}unb.br

Received October 16, 2007; Accepted November 15, 2007


  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 harboring a KEX2 processing site between both chains. After KEX cleavage, a correctly folded mature Fab is formed. The produced recombinant protein was characterized as a heterodimeric functional Fab.The vector described is a new tool for the proper expression of antibody fragments or any heterodimeric polypeptides.

Key Words: Antibody engineering, Pichia pastoris, Fab expression, Kex


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.