https://dx.doi.org/10.1002/jps.24454">
 

Title

Comparative Study of Analytical Techniques for Determining Protein Charge

Abstract

As interest in high-concentration protein formulations has increased, it has become apparent that routine, accurate protein charge measurements are necessary. There are several techniques for charge measurement, and a comparison of the methods is needed. The electrophoretic mobility, effective charge, and Debye-Hückel -Henry charge have been determined for bovine serum albumin, and human serum albumin. Three different electrophoretic methods were used to measure the electrophoretic mobility: capillary electrophoresis, electrophoretic light scattering, and membrane confined electrophoresis. In addition, the effective charge was measured directly using steady-state electrophoresis. Measurements made at different NaCl concentrations, pH, and temperatures allow comparison with previous charge estimates based on electrophoresis, Donnan equilibrium, and pH titration. Similar charge estimates are obtained by all of the methods. The strengths and limitations of each technique are discussed, as are some general considerations about protein charge and charge determination.

Department

Molecular, Cellular and Biomedical Sciences

Publication Date

7-1-2015

Journal Title

Journal of Pharmaceutical Sciences

Publisher

Elsevier

Digital Object Identifier (DOI)

https://dx.doi.org/10.1002/jps.24454

Document Type

Article

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