Ueda T, Maeda Y, So T, Imoto T
Graduate School of Pharmaceutical Sciences, Kyushu University 62, Fukuoka, Japan.
Cell Mol Life Sci. 1997 Dec;53(11-12):929-34. doi: 10.1007/s000180050113.
Recently we developed a slow dialysis method that effectively refolds denatured and reduced immunoglobulin G (IgG) [Maeda, Ueda and Imoto (1996) Prot. Engng 9: 95-100]. This method allows both individual and simultaneous refolding of denatured and reduced H and L chains. Analysis by SDS-polyacrylamide gel electrophoresis revealed that some oligomers were formed through disulfide bonds when H chains were refolded individually. It was also shown that the extent of IgG obtained by rejoining the mixture of refolded H and L chains which had been refolded individually was similar to that obtained by refolding denatured and reduced whole IgG. The results indicated that a favourable interaction between H and L chains prevented formation of H-chain oligomers to yield intact IgG. The present results suggest a mechanism whereby individually folded chains might associate to form IgG molecules in vivo.
最近我们开发了一种缓慢透析方法，该方法能有效地使变性和还原的免疫球蛋白G（IgG）重新折叠[前田、上田和井本（1996年）《蛋白质工程》9：95 - 100]。这种方法允许变性和还原的重链和轻链单独或同时进行重折叠。通过SDS - 聚丙烯酰胺凝胶电泳分析表明，当重链单独重折叠时，一些寡聚体通过二硫键形成。还表明，将单独重折叠的重链和轻链混合物重新结合所获得的IgG程度与变性和还原的完整IgG重折叠所获得的程度相似。结果表明，重链和轻链之间的有利相互作用可防止重链寡聚体的形成，从而产生完整的IgG。目前的结果提示了一种机制，即单独折叠的链可能在体内缔合形成IgG分子。
