I'm trying to get the soluble expression of Fab(Fragment of Antibody) in cytoplasma of E.coli.I have gone through the no of litratures and tried some experiment but not getting enough expression.
Actually my question is that ,What we can do as a upstream member (fermentation) to get the good level of soluble expression in e.coli?
I have tried lower temprature,lower agitation,low level of inducer concentration and used some modified media also to get the soluble expression but still not getting enough expression levels.
Please give your suggestions.
To achieve high level expression of recombinant protein in large scale, I recommend you try pilot expression optimization at first, among which coden optimization may be the key piont. Codon optimization can produce pauses to slow down translation rate and balance the essential factors of soluble protein expression. By optimization of various factors such as codon usage bias, GC content and mRNA structure, we can solve the problems of soluble protein expression and protein yield improvement. Furthermore, high cell density fermentation by fed-batch strategies is one of the most cost-effective means of achieving high yields for the production of large scale recombinant proteins.