coupling peptides

rmsmith at unix.amherst.edu rmsmith at unix.amherst.edu
Sun Jan 8 09:51:39 EST 1995


Hallick Lab (hallick_lab at TIKAL.BIOSCI.ARIZONA.EDU) wrote:
: 	I have a question about coupling peptides to carrier proteins for use in
: immunizations.  I am interested in using EDC to couple my peptides to BSA.  I
: would like to know people's opinion of this method.  Is it tricky?  Is it a
: fairly successful method?  I am worried about blocking free amino groups on
: the peptide.  Is this necessary?  (Each of my 10mer peptides has at least two
: internal lysines.)  I would appreciate any hints or comments.
: Thanks,
: Kristin Jenkins
: Dept. of Molecular and Cellular Biology
: University of Arizona
: hallicklab at tikal.biosci.arizona.edu

   We do this kind of thing in our lab all the time.  While I have never
done this I believe the method we use involves activating the epsilon
amino groups on protein lysines with Trauts reagent to yield more
reactive thiol groups.  We then use a heterobifunctional crosslinker (a
maleomidocaproate or something like that) to react first with the
peptide.  Your unprotected peptide lysines would be a problem here in
our scheme but you might be able to find a way around this such as to
couple via the C-terminus.  The Trauts reagent strategy along with a
plethora of crosslinking reagents can be found in the Pierce catalog
(with a few references).  If you want procedures or any other
information feel free to email me.

Gook Luck.

Rob.

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
    Robert M Smith                        rmsmith at amhux3.amherst.edu

Department of Chemistry,     a      Program in Molecular and Cellular Biology
    Amherst College,           n          University of Massachusetts,
   Amherst, MA 01002             d             Amherst, MA 01003

 Phone:  (413) 542-2558                      FAX:  (413) 542-2735 
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~





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