scFv purification- how ?

Svend Kjaer kjaer at biobase.dk
Wed Nov 15 06:45:27 EST 1995


Hi there !

We are working with the Nissim library from Greg Winthers lab, MRC. We have
found pretty good antibodies against a variety of antigens. The antibody-
fragments look specific when tested in western blots using the scFv on the
phage. However, for a number of purposes, it is more convenient to have
the scFv by itself. The production of these fragments is rather straighfor-
ward, simply moving the phagemid with the scFv insert to a non-suppressor
strain like HB2151 and then grow it and induce by IPTG.
We have gotten so far, but are stalling a bit on the purification of
large amounts of the expressed scFv,i.e. from 1-2 liters cultures.

So I would like to hear from anyone with such purification experiences,
what do you do ? , strains, induction periods, fractionating, columns etc.

Thanks

Svend



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