Hi Neil,
I'm no peptide chemist, but I collaborated with a very sharp one a few
years ago. Here's her protocol:
The reaction is just the same as an AA coupling. The N-terminal amine of
the protected peptide resin is blocked by treatment with biotin
N-hydroxysuccinimide ester (4 equiv), HOBt (4 equiv), and DIEA (8 equiv) in
DMF for 16 hr at 25 degrees C. Check by ninhydrin if desired. The
biotinylation reagent may not be soluble at first, but dissolves as the
reaction proceeds.
Then cleave the resin.
Dissolving the crude biotinylated peptides in water may require adjusting
to neutrality.
Biotin N-hydroxysuccinimide can be (could be?) purchased from Sigma or
Fluka. Alternatives to give a variety of spacers between the biotin and
the peptide should also be available.
'luck!
Steve Hartson
-------
>Dear All
> I have got to make some peptides labelled with biotin and
>fluorescein. This will be the first time I have attmpted such labelling and
>I wonder if anybody out there in ABRF-land has any Protocols/references for
>on resin labelling of peptides with such labels as biotin/fluorescein/DIG
>etc.
>
> Many thanks in advance, Neil Brew
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