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.
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The Carlsberg Foundation is gratefully acknowledged for financial support.
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Højrup, P. (2015). Analysis of Peptides and Conjugates by Amino Acid Analysis. In: Houen, G. (eds) Peptide Antibodies. Methods in Molecular Biology, vol 1348. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2999-3_8
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DOI: https://doi.org/10.1007/978-1-4939-2999-3_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2998-6
Online ISBN: 978-1-4939-2999-3
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