Working on the DG44 CHO cells for recombinant protein expression, we have been getting the low expression of 0.5ug/ml recombinant protein and also Gal-a(1,3)Gal glycosylation of protein. Glycosylation pattern has to be improved to Gal-a(2,3/6)Gal + improvement of expression to mg/ml level. Suggestion and collaboration are welcome.
Thanks and regards,
G IRISH
Gannareddy: There are millions of factors affecting the yield and glycosylation pattern of a recombinant protein in CHO expression system. For example, we intended to overexpress a lysosomal glycoprotein enzyme and expected to obtain a uniformed high-mannose containing phosphate (Man-6-P) as the natural intracellular protein, but instead getting a protein in the medium with a mixture of all kinds of carbohydrate structures including Man-6-P, which severely reduced its usefulness as a therapeutic agent. So, think about the natural protein and its targeting route, and choose a right expression system (CHO or its mutants, or other mammalian cells). Good luck.
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