I am trying to express the Fab in E. coli. I am using a duet vector with T7 promoter system for expression. Unfortunately there is an unequal expression of light and heavy chain. Any insights into how I can equally express the light and heavy chain?
Hi,
Look at this paper:
http://www.sciencedirect.com/science/article/pii/S1046592802005430
A plasmid system for optimization of Fab′ production in Escherichia coli: importance of balance of heavy chain and light chain synthesis
Protein Expression and Purification
Volume 26, Issue 2, November 2002, Pages 309–320
'The LC:HC balance has been controlled by varying the codon usage of the signal peptide (SP) and 5′ mature domain coding regions.'
Good luck!
Rare codon optimization. The possible reason is that the gene for one chain contain contain more rare codons than another chain, So one better way is to optimize the rare codons for both chains.
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