antibodies from inclusion bodies

D.K. at
Wed Apr 23 16:39:59 EST 2003

In article <20030423185347.12208.qmail at>, humphreyt at wrote:
>My recombinant protein is strongly expressed in inclusion bodies in E coli.  I
> have heard that purified inclusion bodies can be directly injected into
> rabbits for antibody production without the need for solubilising and
> refolding.  Has anyone tried this and / or have any hints and tips for me?

Yeah, it's been tried many times. It's quick and dirty. Invariably,
such sera work but show a lot of cross-reactivity/high background.
Considering how easy it is to purify 6His-tagged protein under 
denaturing conditions (or even non-tagged by ion exchange in the 
presence of urea) and use it in immunizations, I personally 
find there is no excuse for using crude inclusion preps in 
immunizations. No refolding necessary - you can inject unfolded 
precipitated protein.


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