Peptide conjugation efficiency
jeffrey_weidner at merck.com
Wed Nov 1 12:36:14 EST 1995
In article <46mju1$ock at ixnews7.ix.netcom.com>, nancied at ix.netcom.com
(Nanci E Donacki ) wrote:
> Does any one know of a procedure to determine the efficiency of a
> peptide coupled to a protein carrier? The only method I know of
> requires radiolabeled peptides, and then these can't be used for
> immunization of animals.
> Are there any other procedures that can be used?
> Thanks in advance.
> Nanci E Donacki
If you incorporate a non-native amino acid such as norleucine into your
peptides, you can easily determine coupling efficiency by performing an
acid hydrolysis and amino acid analysis on your coupled protein. We
routinely do this to monitor our coupling efficiency and we generally
prepare 100-150 peptide immunogens per year. The Nle should be placed as
clsoe to the cross-linking moiety as possible in order to minimize it's
contribution to the antigenicity of the desired peptide sequence.
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