Improving SDS-PAGE method for monoclonal antibodies: The advantages of Tris-Acetate over Tris-Glycine SDS-PAGE system and comparison with CE-SDS method
| Autor: | Mohsen Yazdani, Rozhan Sonboli, Hossein Behrouz, Zeinab Najafi, Nahid Zarezadeh |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Tris 0303 health sciences Chromatography Resolution (mass spectrometry) medicine.drug_class Chemistry Glycine Antibodies Monoclonal Acetates Monoclonal antibody 01 natural sciences 03 medical and health sciences Electrophoresis chemistry.chemical_compound Capillary electrophoresis 010608 biotechnology medicine Electrophoresis Polyacrylamide Gel Tromethamine Polyacrylamide gel electrophoresis 030304 developmental biology Biotechnology |
| Zdroj: | Protein Expression and Purification. 182:105845 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2021.105845 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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