Analysis of Peptides and Conjugates by Amino Acid Analysis

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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.

Original languageEnglish
Title of host publicationPeptide Antibodies : Methods and Protocols
EditorsGunnar Houen
PublisherHumana Press
Publication date2015
Pages65-76
ISBN (Print)978-1-4939-2998-6
ISBN (Electronic)978-1-4939-2999-3
DOIs
Publication statusPublished - 2015
SeriesMethods in Molecular Biology
Volume1348
ISSN1064-3745

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Amino Acids
Peptides
Ninhydrin
Ion Exchange Chromatography
Glutamine
Amides
Tryptophan
Glutamic Acid
Hydrolysis
Gases

Cite this

Højrup, P. (2015). Analysis of Peptides and Conjugates by Amino Acid Analysis. In G. Houen (Ed.), Peptide Antibodies: Methods and Protocols (pp. 65-76). Humana Press. Methods in Molecular Biology, Vol.. 1348 https://doi.org/10.1007/978-1-4939-2999-3_8
Højrup, Peter. / Analysis of Peptides and Conjugates by Amino Acid Analysis. Peptide Antibodies: Methods and Protocols. editor / Gunnar Houen. Humana Press, 2015. pp. 65-76 (Methods in Molecular Biology, Vol. 1348).
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Højrup, P 2015, Analysis of Peptides and Conjugates by Amino Acid Analysis. in G Houen (ed.), Peptide Antibodies: Methods and Protocols. Humana Press, Methods in Molecular Biology, vol. 1348, pp. 65-76. https://doi.org/10.1007/978-1-4939-2999-3_8

Analysis of Peptides and Conjugates by Amino Acid Analysis. / Højrup, Peter.

Peptide Antibodies: Methods and Protocols. ed. / Gunnar Houen. Humana Press, 2015. p. 65-76 (Methods in Molecular Biology, Vol. 1348).

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

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Højrup P. Analysis of Peptides and Conjugates by Amino Acid Analysis. In Houen G, editor, Peptide Antibodies: Methods and Protocols. Humana Press. 2015. p. 65-76. (Methods in Molecular Biology, Vol. 1348). https://doi.org/10.1007/978-1-4939-2999-3_8