The present invention provides a method for producing a multimeric protein composed of a monomeric protein, wherein the monomeric protein is obtained by fusing a protein having an immunoglobulin fold structure to a protein that can serve as a subunit structure, the method including the steps of:
(A) preparing the monomeric protein having an insoluble granular form in cells of a microorganism;
(B) solubilizing the monomeric protein prepared in step (A) with an aqueous solution containing lauroyl-L-Glu;
(C) diluting a solution obtained in step (B) in a buffer containing arginine hydrochloride to lower a concentration of lauroyl-L-Glu; and
(D) replacing a solvent of a solution obtained in step (C) with a buffer using gel filtration chromatography or the like.
本发明提供了一种生产由单体蛋白质组成的多聚体蛋白质的方法,其中单体蛋白质是通过将具有免疫球蛋白折叠结构的蛋白质与可作为亚单位结构的蛋白质融合而得到的,该方法包括以下步骤:
(A) 在微
生物细胞中制备具有不溶性颗粒状的单体蛋白;
(B) 用含有月桂酰-L-Glu 的
水溶液增溶步骤(A)中制备的单体蛋白;
(C) 在含有
盐酸精
氨酸的缓冲液中稀释步骤(B)中得到的溶液,以降低月桂酰-L-谷
氨酰的浓度;以及
(D) 使用凝胶过滤色谱法或类似方法将步骤(C)中得到的溶液的溶剂替换为缓冲液。