covalent coupling strategies

Roger Murphy murphy_r at licre.ludwig.edu.au
Mon Dec 1 00:57:21 EST 1997

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,
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

Tel  61-3-94965463
Fax  61-3-94965436
Email murphy_r at licre.ludwig.edu.au

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