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    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. Copyright © 2021 Elsevier Inc. All rights reserved.

    Citation

    Rozhan Sonboli, Zeinab Najafi, Nahid Zarezadeh, Mohsen Yazdani, Hossein Behrouz. Improving SDS-PAGE method for monoclonal antibodies: The advantages of Tris-Acetate over Tris-Glycine SDS-PAGE system and comparison with CE-SDS method. Protein expression and purification. 2021 Jun;182:105845

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    PMID: 33596474

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