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P. Minkiewicz, J. Dziuba, M. Niklewicz

RAPID COMMUNICATION
Protein homogeneity testing by reversed-phase high-performance liquid chromatography

of reduced and non-reduced samples (in English)

Polimery 2005, No 5, 379




Summary

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 α-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.


Key words: disulphide bonds, α-lactalbumin, protein standard heterogeneity, reversed-phase high-performance liquid chromatography


P. Minkiewicz, J. Dziuba, M. Niklewicz

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