Toward more efficient protein expression: keep the message simple.

Abstract:

:Optimization of gene coding-sequence, including preferred codon usage and removal of cryptic splice sites and mRNA-destabilizing motifs, has been shown to improve recombinant protein production of different proteins. Here, we present data to show that gene optimization can also be used to improve the production of a complex macromolecule, namely an antibody. When applied to the heavy and light chain genes of our model antibody, we found that greater numbers of high-producing transfectants as well as increased levels of protein production were observed (approximately 1.5-fold). In this test model, production was improved even though the antibody has previously been demonstrated to give high expression in stably transfected cells (up to 5 g/L in bioreactors). Because the parental heavy chain sequence contained introns, and the process of gene optimization is most efficiently performed on sequences without introns, we demonstrated that removal of introns in the coding sequence had no effect on the quantity of antibody produced. All constructs were evaluated using Lonza's glutamine synthetase gene expression vectors in Chinese hamster ovary cells. Our findings suggest that significant improvements in product yields can be achieved by gene optimization, which may facilitate the processing and translation of gene transcripts.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Kalwy S,Rance J,Young R

doi

10.1385/MB:34:2:151

subject

Has Abstract

pub_date

2006-10-01 00:00:00

pages

151-6

issue

2

eissn

1073-6085

issn

1559-0305

pii

MB:34:2:151

journal_volume

34

pub_type

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