I have started using Invitrogen's freestyle CHO cells (without serum, and in suspension) but the protein expression levels are not great compared to the HEKs I was using before.
I have played around with ratios and have an optimal ratio (1:1 with 1ug DNA and 1ug MAX transfection reagent). I have also tried putting the cells in hypothermic conditions which appears to have increased the expression by almost double, but this still isn't quite enough.
Any other suggestions of something I can do (without extra cloning) to imrpove the expression of the cells?
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#2
genehunter
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Posted 19 August 2009 - 09:27 AM
Its hard to beat the transfection rate of 293.
Have you done any EGFP or beta-Gal transfection to test what % of transfection do you have?
Have you done any EGFP or beta-Gal transfection to test what % of transfection do you have?
#3
niki
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Posted 20 August 2009 - 02:10 AM
It is hard to beat it I know, we had the wrong glycosylation patterns though. Just want to up the production as much as possible.
We transfected with GFP, gave about a 60% transfection rate.
Anyone who has any expreience with mammalian cells can help though, I'm not just looking for tricks specific to CHOs. I will try anything that has been shown to work on mammalian cells.
We transfected with GFP, gave about a 60% transfection rate.
Anyone who has any expreience with mammalian cells can help though, I'm not just looking for tricks specific to CHOs. I will try anything that has been shown to work on mammalian cells.
