Analysis of Peptides and Conjugates by Amino Acid Analysis

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Abstract

Amino acid analysis is a highly accurate method for characterization of the composition of synthetic peptides. Together with mass spectrometry, it gives a reliable control of peptide quality and quantity before conjugation and immunization.Peptides are hydrolyzed, preferably in gas phase, with 6 M HCl at 110 °C for 20-24 h and the resulting amino acids analyzed by ion-exchange chromatography with post-column ninhydrin derivatization. Depending on the hydrolysis conditions, tryptophan is destroyed, and cysteine also, unless derivatized, and the amides, glutamine and asparagine, are deamidated to glutamic acid and aspartic acid, respectively. Three different ways of calculating results are suggested, and taking the above limitations into account, a quantitation better than 5 % can usually be obtained.

OriginalsprogEngelsk
TitelPeptide Antibodies : Methods and Protocols
RedaktørerGunnar Houen
ForlagHumana Press
Publikationsdato2015
Sider65-76
ISBN (Trykt)978-1-4939-2998-6
ISBN (Elektronisk)978-1-4939-2999-3
DOI
StatusUdgivet - 2015
NavnMethods in Molecular Biology
Vol/bind1348
ISSN1064-3745

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