DESIGN OF PEPTIDE?

altabios at bham.ac.uk altabios at bham.ac.uk
Fri Mar 24 10:49:58 EST 2000



Tilman Lamparter wrote:

> I would like to order peptides for protein interaction studies. Maybe
> somebody can help me with the following questions: There are two 8mer
> (VAIRQYCC and CFILCNSC) peptides we want to test for binding to a protein.
> The sequence information was given by a phage display. Binding assay
> should either be done with immobilized peptide, coupled to Affigel 10 or
> 15 (Biorad) or with biotinilated peptide, which later should bind to a
> streptavidin matrix. I think in both cases, a linker is needed to leave
> space between the matrix and the peptide. How should this be designed? If
> coupling to affigel (via amino end or side chain of peptide and
> succinimido-groups of the gel) should be efficient, is it correct/ better
> to add lysin at one end (to increase the yield) ?
>

I have just looked at your message in a bit more detail.
We can make peptides coupled to controlled pore glass, it simply avoids having
to try to put them back onto a solid phase again.
Typically we use 1000A CPG at a loading of about 20umole/g, great for affinity
columns.
The link is at the C-terminal of the peptide.

We have no hard information about spacers but could easily add 2 glycines
between the peptide and the linker.

John Fox

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