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Online capillary electrophoresis – mass spectrometry analysis of histatin-5 and its degradation products

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Abstract

Histatin-5 (Hst-5) is a human salivary peptide with antibacterial and antifungal activities. Thorough characterization and reliable quantification of Hst-5 and its degradation products are essential for understanding the Hst-5 degradation pathway. Due to the highly basic and strong cationic nature of the Hst-5 peptide, the quantitative analysis of Hst-5 and its degradation forms by online mass spectrometry remains challenging. Here, we adopt a recently developed electrokinetically pumped sheath liquid capillary electrophoresis – mass spectrometry (CE-MS) coupling technology, and successfully apply it for the analysis of Hst-5 and its degradation products. Our CE-MS method is demonstrated to be robust and quantitative. This novel analytical platform is reproducible and free of sample carryover. The efficacy of this method is demonstrated with a kinetic study of Hst-5 degradation by Sap9, a secreted aspartic peptidase. Our work demonstrates the potential of online CE-MS as a powerful approach for characterizing highly basic peptides.

Graphical abstract: Online capillary electrophoresis – mass spectrometry analysis of histatin-5 and its degradation products

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Article information


Submitted
15 Apr 2020
Accepted
09 Jun 2020
First published
11 Jun 2020

Analyst, 2020, Advance Article
Article type
Paper

Online capillary electrophoresis – mass spectrometry analysis of histatin-5 and its degradation products

J. Lamp, S. P. Ikonomova, A. J. Karlsson, Q. Xia and Y. Wang, Analyst, 2020, Advance Article , DOI: 10.1039/D0AN00756K

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