peptide biotinylation question

Austin P. So (Hae-Jin) haejin at netinfo.ubc.caX
Tue Nov 14 13:49:09 EST 2000


Reverse phase HPLC is your best bet...no ambiguity.

You may run into the same problems with your dialysis membrane wrto MW
cutoff...depends if there is a suitable membrane available.

If you want to play around a bit, you could try selectively precipitating the
Biotin-X-OH (the NHS ester is hydrolysed) through weak acid treatment (0.01N
HCl), since biotin-X-OH is pretty insoluble in aqueous solution...really depends
on what coupling conditions you used.

Student wrote:

> I need to biotinylate two 10-amino acid peptides with NHS-X-Biotin. My
> problem is this: how can I effectively remove all the unreacted free biotin
> ester after the biotinylation reaction? I can't use the normal desalting
> methods because the biotinylated peptide is still too small.

--
---
Austin P. So (Hae Jin)

I.I.S.G.P.
Biotechnology Laboratory
University of British Columbia

E-mail: haejin at netinfo.ubc.ca

http://www.interchange.ubc.ca/haejin/index.html (under construction)







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