Dear Alessandro,
after long years working with peptides, I am sorry to say that amino acid
analysis is the only reliable method, in my eyes. The drawbacks beside the
work are either amino acids which are destroyed or amino acids which are
generally to high, but in case of a known sequence - this should be with a pure
synthetic peptide - you can look for the most stable amino acids. There is
still a small error but less than any other method. I used 160 ƒC vapour
phase hydrolysation and OPA/FmoC precolumn derivatisation on a HP Aminoquant
with RP separation and either UV or fluoresecence detection, standard
protocolls. In comparison to other methods (Ninhydrin etc.) the results were within
small deviations, even if the whole procedure had been carried out in a
different lab on the same peptide.
Yours
Manfred Raida
APB Proteomics
-- Dr. Manfred Raida European Proteomics Lab Amersham Pharmacia Biotech Munzingerstrasse 9 D-79111 Freiburg Germany Email: Manfred.Raida@eu.apbiotech.com
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