Tobias Dassler wrote:
>> I'm working with a membrane protein of E.coli which is quite hydrophobic
> and which I can't overproduce in sufficient amounts for purification and
> antibody production. So I'm trying to find special peptide-sequences of
> the protein for use as antigens.
> Now my questions:
> Does anyone out there know anything about the properties such a peptide
> sequence has to have to be suitable as an antigen? Or which amino acids
> shouldn'd be present at all? Are there any algorithms for calculating the
> "antigenicity" of a peptide? Are there any servers/programs in the net to
> get informations from?
> Thanks for your help!
> Tobias
The only criteria is that for the peptide sequence selected to be of
real use, is that it should be from the protein surface. Obviously to
allow the use of the subsequent antibodies, to this key sequence, in
work with the intact, native conformation, parent protein. So clearly a
relatively hydrophillic region would be a good starting point. Peptides
for antibody production are typically 8-12 reisdues. So you might want
to try a couple of sequences and see which gives you best selectivity
for your native parent protein.
The more promenient the sequence motif is on the protein surface the
more readily it will be identified. Typically the target peptide is
conjugated to a carrier protein, say KLH specificaly by one end of the
peptide, for antibody production so an immune response is almost assured
in mammals.
Im not aware of any formal protocols for calculating such things as
'antigenicity' or for identifying appropriate protein segments for use,
it seems to basically comes down to experience and common sense.
I hope this helps, Regards,
Len.
--
Dr Len Bell,
University of Liverpool.
email: lgbell at liv.ac.uk
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