The Effect of Temperature in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis of Protein Therapeutics

Filep, Csenge and Guttman, András (2020) The Effect of Temperature in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis of Protein Therapeutics. ANALYTICAL CHEMISTRY, 92 (5). pp. 4023-4028. ISSN 0003-2700

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Abstract

The temperature-dependent migration of molecular weight protein size standards and several biotherapeutic proteins were studied in sodium dodecyl sulfate capillary gel electrophoresis (SDS-CGE) in the interval from 15 to 60 °C using borate crosslinked dextran sieving matrix. Arrhenius plots were generated to calculate the respective activation energy values for the various solute molecules. SDS-CGE analysis of the biotherapeutic protein test mixture revealed no correlation between the activation energy requirement of the different species and their molecular weights, emphasizing the importance of separation temperature optimization to obtain high resolution between the solute molecules of interest. In contrast, the molecular weight protein size ladder ranging from 10 to 225 kDa, built from the same polypeptide blocks with no posttranslational and other modifications, showed predictable activation energy requirement. The electrophoretic mobility of the SDS−protein complexes was found to be the function of the reciprocal sixth root of the molecular weight (Mw −1/6), implying cylindrical conformation.

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