Protease inhibitors

[NANTEL at UNCVX1.BITNET]

Greetings Netters!!

I'm fishing for information concerning the use of protease inhibitors during
the purification of MBP-fusion proteins from E.coli. PMSF doesn't seem to
have any effects on the quality of my protein preps. Furthermore, when I
use antipain + leupeptin the efficiency of the purification step through
the amylose column is greatly reduced (7-10 fold lower yields). I feel
that protease inhibitors are necessary since one of my fusion protein is
chewed up from 95 to 80 kD during the purification.

I am faily new to the world of protein purification (yes, another molecular
biologist doing biochemistry without a licence) and any suggestions would
be most welcome.

Andre Nantel
Biology Dept.
University of North Carolina