Biopharmaceutical Research Center, AryoGen Pharmed Inc., Alborz University of Medical Sciences, Karaj, Iran.
Biopharmaceutical Research Center, AryoGen Pharmed Inc., Alborz University of Medical Sciences, Karaj, Iran.
Protein Expr Purif. 2021 Jun;182:105845. doi: 10.1016/j.pep.2021.105845. Epub 2021 Feb 14.
Present study compares two different buffer systems for the electrophoretic separation of the IgG and IgG 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.
本研究比较了两种不同的缓冲体系，用于 SDS-PAGE 方法电泳分离 IgG 和 IgG 单克隆抗体。与传统使用的 Tris-甘氨酸方法相比，改良的 Tris-乙酸盐系统在 6-20%梯度凝胶中分离这些蛋白质具有优越性。与传统的 Tris-甘氨酸方法相比，这种改良的 Tris-乙酸盐缓冲系统显示出更清晰的条带、更准确的分子量测定、更高的分辨率和更好的亚片段估计，与毛细管凝胶电泳获得的结果更接近。在平行实验中，还研究了 PNGase-F 酶对 IgG 去糖基化的影响，结果表明对 SDS-PAGE 结果没有显著改善。
