Protein homogeneity testing by reversed-phase high-performance liquid chromatography of reduced and non-reduced samples

P. Minkiewicz; J. Dziuba; M. Niklewicz

Published : 2022-09-01

Vol. 50 No. 5 (2005)

Protein homogeneity testing by reversed-phase high-performance liquid chromatography of reduced and non-reduced samples

P. Minkiewicz J. Dziuba M. Niklewicz

Abstract

The concept of this study was based on the assumption that protein standards obtained using the same method may reveal heterogeneity explainable by changes in the disulphide bonding pattern. Two lots of bovine a-lactalbumin standard were subjected to reversed-phase high-performance liquid chromatography (RP-HPLC). One of them was homogenous. The other revealed the presence of two major fractions when non reduced. Neither RP-HPLC after protein reduction nor mass spectrometry showed differences between them. This indicates that one of the lots contained isomers with different locations of disulphide bonds. A comparison of RP-HPLC profiles obtained with and without reduction may be thus recommended as a tool for testing homogeneity of protein standards, especially those used in experiments involving denaturation.

Keywords

wiązania disiarczkowe ; laktoalbumina-a ; heterogeniczność wzorców białek ; wysokosprawna chromatografia cieczowa z odwróconymi fazami disulphide bonds ; a-lactalbumin ; protein standard heterogeneity ; reversed-phase high-performance liquid chromatography

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Minkiewicz, P., Dziuba, J., & Niklewicz, M. (2022). Protein homogeneity testing by reversed-phase high-performance liquid chromatography of reduced and non-reduced samples. Polimery, 50(5), 379–382. Retrieved from https://ichp.vot.pl/index.php/p/article/view/1644

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P. Minkiewicz

Affiliation
University of Warmia and Mazury in Olsztyn, Chair of Food Biochemistry, Plac Cieszyński 1, 10-726 Olsztyn Kortowo, Poland

J. Dziuba jerzy.dziuba@uwm.edu.pl

Affiliation
University of Warmia and Mazury in Olsztyn, Chair of Food Biochemistry, Plac Cieszyński 1, 10-726 Olsztyn Kortowo, Poland

M. Niklewicz

Affiliation
University of Warmia and Mazury in Olsztyn, Chair of Food Biochemistry, Plac Cieszyński 1, 10-726 Olsztyn Kortowo, Poland