I am wondering what people's experiences are with protein expression in Sf9/Sf21 cells versus Hi5's? Are Hi5's usually better, is it work screening expression of all targets in Sf's and Hi5's? Thanks!
My experience is with Sf9 and Hi5 cells. We use Sf9 for all routine work (transfection, virus amplification) and we screen expression of most targets in both cell lines. Both cell lines behave well in 24 deep well blocks and shake flasks. In general, we have higher secretion yield with Hi5 than Sf9. However, there are exceptions. For intracellular proteins, I don't observe one better than the other.
Cell line also affects downstream such as protein purification. For example, protein purity of the target using the same chromatography technique varies between the cell lines. Glycosylation is also different.
Thanks for your feedback!
we use SF9 for transfection and BIICs
For expression we always test both cell lines, SF9 and H5. H5 tend to produce 5x more than SF9 and grow faster. But they also tend to show more proteolysis than SF9. Thats why we first enrich from a small scale, do LCMS and only if mass is correct go with that for production.
However, if the western blot comparison of both cell lines clearly show degradation in H5 and none in SF9, we skip H5.