Pichia recombinant proteins
bspinoza at aol.com
Thu Jul 6 20:02:17 EST 1995
I have been studying the human erythropoietin receptor
extracellular domain as a bacterially expressed GST fusion protein. I
became unsatisfied with the activity of this recombinant protein- only 1%
of the soluble protein bound ligand with the expected high affinity, even
after tedious affinity purification. Consequently, we have expressed
this protein in the yeast Pichia pastoris. It seems to have a much higher
specific activity than the bacterially expressed protein (still working on
how much...) and is appropriately glycosylated (by molecular weight).
Unfortunately the yields are, to say the least, disappointing.
I have three questions:
1. Are a lot of people using this system (Invitrogen)?
2. What is the experience with improving yields?
3. Anyone have any ideas for an interesting collaboration with this
Kevin Harris- harrisk at uthscsa.edu
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