Soluble expression

Benny benny at holmes.nott.ac.uk
Tue Nov 21 17:35:24 EST 2000


Have you included a pelB signal before the antibody-encoding DNA to direct
any translated protein to the periplasm? In addition to this, inducing E
coli at lower temperatures may sometimes help. The reason I said "sometimes"
is because whether soluble protein or inclusion bodies are produced is very
sequence-dependent and varies from one antibody to another.

If you are making variable region fragments, have a look at APBiotech's
recombinant phage antibody system. We have used them and, although the
yields are low, soluble antibody fragments are detected at the supernatant
and the periplasmic extract. It contains everything you need from gene
construction to protein expression.

Hope this helps.

Benny.


"Richard Buick" <Richard.Buick at btinternet.com> wrote in message
news:8velia$2ut$1 at uranium.btinternet.com...
> Does anyone know of any conditions for expressing soluble antibody
fragments
> in E. coli? All my attempts so far have produced either inclusion bodies
or
> nothing at all.
> Thanks
> Rick
>
> --
>
>
>
> ***************************************************
> Richard J. Buick
>
> E-mail: mail at richardbuick.com
> URL: http://www.richardbuick.com
> Tel(GSM): +44 7801 416792
>
> ***************************************************
>
>







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