Antibodies against inclusion body proteins?

Simon Dawson Simon.Dawson at nott.ac.uk
Mon Mar 18 11:00:12 EST 1996


Chris Jones wrote:
> 
> Hello,
>            We have the age old problem of our E.coli expression
> construct being insoluble and in inclusion bodies. As we only wish to
> prepare Ab rather than demonstrate activity do we really need a fully
> refolded and soluble protein for rainsing Ab? Actually our expression
> construct is a His Tag PCR fragment of part of a coding region so we
> can't expect to get Ab to the 3d form of the full length protein. I am
> worried that injecting what is essentially an insoluble protein will
> fail to give an antigenic response. Any suggestions???
> 
> Thanks
> 
> Chris J

Hi,
   Im my experience (which isn't a lot I admit!), insoluble antigens used in 
raising polyclonal antisera often work extremely well. Better than soluble forms 
of the same protein in some cases. The only reason that I can think of is that 
the insoluble antigen is cleared from the animals system slower than one which is 
soluble and so can stay around longer to elicit an immune response.
  Hope thats of some help,


         Simon.

-- 

Dr. Simon Dawson       TEL:+44 (0)115 9249924 ex. 44787
Dept. of Biochemistry  FAX:+44 (0)115 9422225
Queens Medical Centre  Email:Simon.Dawson at .nott.ac.uk
Clifton Boulevard      http://www.ccc.nottingham.ac.uk/~mbzspd/Simon.html
Nottingham
U.K.                   "Back off man, I'm a scientist!" - Bill Murray.



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