MAP peptides for Abs
bernard at elsie.nci.nih.gov
Fri Apr 29 01:42:32 EST 1994
In article <2pp1uk$c7 at u.cc.utah.edu>, Tom Doak <tom_doak at hlthsci.med.utah.edu> writes:
> We hope to make antisera to synthetic peptides representing our favorite
> ORF. I had thought to do this by a fairly conventional route: have the
> pep synthesised, conj to KLH with a Pierce kit, have it injected into
> rabbits, and hope. However the AppliedBiosystems MAP substrate for
> peptide synthesis has been suggested, but not by anyone with much
> experience using it. So MAPs are Multiply Attacted Peptides, or some
> such, giving a small tree of peptides, each branch is another of the
[Some stuff deleted for brevity]
> Thanks so very much,
> Tom Doak
> tom_doak at hlthsci.med.utah.edu
My only feeling is that you can't predict how good the MAP approach
will be for your particular peptide. As an immunogen it is certainly not as
reliable as KLH. We tried it for one of our peptides and found no antibody,
yet the same peptide coupled to KLH gave great antibodies, and those antibodies
bound very well to the MAP in ELISA (they do stick to ELISA plates). Other
hearsay claims that titres are not as good with the MAP as KLH conjugates.
However, other people swear by this technology and its a great approach if
you really don't want anti-carrier antibodies ('though these are easily
removed by back absorption).
So, if you are in a hurry then I'd stick with KLH for the reliability
(I suspect that this is also better for problem peptides - eg. those that
are hydrophobic). If you have the time and money then I'd give the MAP a
whirl as well.
Just my dha/ phingin worth,
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
More information about the Methods