Improving SDS-PAGE method for monoclonal antibodies: The advantages of Tris-Acetate over Tris-Glycine SDS-PAGE system

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Rozhan Sonboli1, Zeinab Najafi1, Nahid Zarezadeh1

  • 1Biopharmaceutical Research Center, AryoGen Pharmed Inc., Alborz University of Medical Sciences, Karaj, Iran.

Protein Expression and Purification
|February 17, 2021
PubMed

Present study compares two different buffer systems for the electrophoretic separation of the IgG1 and IgG2 Monoclonal Antibodies using SDS-PAGE method. A modified Tris-acetate system was shown to be superior for separation of these proteins in a 6-20% gradient gel as compared with the traditionally used Tris-glycine method. This modified Tris-acetate buffer system showed sharper bands, more accurate determination of molecular weight, higher resolution, and better estimation of sub-fragments with closer results to those obtained by Capillary Gel Electrophoresis. Also in a parallel experiment, effect of IgG deglycosylation by PNGase-F enzyme was investigated and revealed no significant improvement on the SDS-PAGE results.

Keywords:
Monoclonal AntibodySDS-PAGETris-AcetateTris-Glycine

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