Maeda Y, Ueda T, Imoto T
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Protein Eng. 1996 Jan;9(1):95-100. doi: 10.1093/protein/9.1.95.
IgG is an oligomeric protein that consists of two heavy and two light chains. To form the oligomer, a highly concentrated protein would be required on renaturation. On the other hand, refolding of proteins at high concentration often led to aggregation. Therefore, denatured and reduced oligomeric protein scarcely refolded to the native structure. As was expected, the folding yield of the denatured and reduced IgG was below 5% under the condition employed in rapid dilution. The low folding yield was elucidated to be due to assembly or aggregation. Using a renaturation method previously developed to depress aggregation effectively by means of slow dialysis, the refolding yield of the denatured and reduced IgG at above 1 mg/ml was above 70%. Most of the refolded IgG was identical with the intact material based on analyses by affinity chromatography and SDS-PAGE.
免疫球蛋白G（IgG）是一种由两条重链和两条轻链组成的寡聚蛋白。要形成寡聚物，复性时需要高浓度的蛋白质。另一方面，高浓度蛋白质的重折叠常常导致聚集。因此，变性和还原的寡聚蛋白几乎不会重折叠成天然结构。不出所料，在快速稀释所采用的条件下，变性和还原的IgG的折叠产率低于5%。低折叠产率被认为是由于组装或聚集所致。使用先前开发的通过缓慢透析有效抑制聚集的复性方法，变性和还原的IgG在浓度高于1 mg/ml时的重折叠产率高于70%。基于亲和色谱和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，大多数重折叠的IgG与完整材料相同。
