ScFv-expression in pVIII???

ijiwaru at wheel.dcn.davis.ca.us ijiwaru at wheel.dcn.davis.ca.us
Wed Jul 31 00:21:00 EST 1996


In article <31FF67DA.D63 at mailserv.cuhk.hk>, b574713 at mailserv.cuhk.hk wrote:

> Dear phage-library workers'
>         I would like to know whether ScFv-antibody normally expressed in
pIII protein can 
> be expressed in pVIII protein. If not, are there any ways to increase
ScFv-antibody copy
> number per phage which is about 1-5 copies per phage in the case of pIII
protein? The
> problem I found is the ScFv-Ab phage derived from my hybridoma  binds
only weakly to
> the antigen.

You probably wouldn't have much success with displaying ScFvs on pVIII as
it is much more finicky about what it will tolerate and still be able to
assemble phage.  You might check out:

Greenwood, et.al. (1991) J. Mol. Biol. 220:821-827
Willis, et.al. (1993)  Gene 128:79-83
Greenwood and Perham (1989)  Virology 171:444-452

I tried to display, what we thought were, very conservative epitopes of no
more than six AAs on pVIII and wasn't able to build phage even in the
presence of wild-type pVIII.  YMMV.

Lyle Najita
Plant Pathology
University of California - Davis



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