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 Expr Purif. 2021 Jun:182:105845. doi: 10.1016/j.pep.2021.105845. Epub 2021 Feb 14.

Abstract

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 Antibody; SDS-PAGE; Tris-Acetate; Tris-Glycine.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / chemistry*
  • Antibodies, Monoclonal / analysis*
  • Antibodies, Monoclonal / chemistry
  • Electrophoresis, Polyacrylamide Gel*
  • Glycine / chemistry*
  • Tromethamine / chemistry*

Substances

  • Acetates
  • Antibodies, Monoclonal
  • Tromethamine
  • Glycine