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.
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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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