Effects of the solution environment on the resolution of recombinant human deoxyribonuclease variants in capillary zone electrophoresis

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作者
Quan, C.P. [1 ]
Canova-Davis, E. [1 ]
Chen, A.B. [2 ]
Chen, Anthony B. [2 ]
Nashabeh, Wassim [2 ]
Wehr, Timothy [2 ]
机构
[1] Department of Analytical Chemistry, Genentech, Inc., One DNA Way, South San Francisco, CA 94080, United States
[2] Department of Quality Control, Genentech, Inc., One DNA Way, South San Francisco, CA 94080, United States
关键词
Capillary electrophoresis - Positive ions - Recombinant proteins - Titration;
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Capillary zone electrophoresis (CZE) is used to demonstrate the interaction of a protein analyte and the surrounding free solution environment, specifically the effects of hydrogen ion or divalent metal cation titration on the resolution of recombinant human deoxyribonuclease I (rhDNase) variants. Hydrogen ion titration of the CZE buffer solution environment observably alters the surface charges of the protein, leading to measurable changes in the electrophoretic mobility which correlates to the theoretical net average protein charge (Z). Similarly, divalent metal cations, when added to the CZE buffer solution, associate with the acidic rhDNase surface charges and lead to observable changes in electrophoretic mobility. Typically, conditions that led to decreased electrophoretic mobility of the protein also led to enhanced resolution of broad zones associated with the heterogeneity of the protein. Additional characterization of the observed heterogeneity distinguished between a distribution of associated ions and glycosylation variation as the source of the protein-associated heterogeneity.
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