Rod,
How about using one of the water-soluble carbodiimide coupling agents (check
out the Pierce catalogue - no commercial connection). I've used these very
effectively. Pre-treat your protein with the carbodiimide for a few minutes to
activate carboxylic acid side chains, then add your peptide which should link
through free primary amino groups (eg Lysine side chains) and leave at 4'C
o/n. Dialyse away the excess eptide and urea by-products and characterize
your conjugate however you wish.
Hope this helps,
Roger
In article <347e058b.1619628 at news.interconnect.com.au>, rmacfarl at csl.com.au
(Rod Macfarlan) wrote:
>I am looking for methods to couple synthetic peptides to a carrier
>protein, ideally through lysine side chains. My problem is that the
>protein is in 8M urea. Is there a way of doing this, and if not, does
>anyone have a suggestion for an alternative approach (no thiols
>available - but thiolation might be possible).
>>Rod Macfarlan
Roger Murphy, Ph.D.
Biological Production Facility
Ludwig Institute for Cancer Research
Austin & Repatriation Medical Centre
Studley Road,
Heidelberg, Vic. 3084
Australia.
Tel 61-3-94965463
Fax 61-3-94965436
Email murphy_r at licre.ludwig.edu.au
