公开(公告)号：US10308969B2

公开(公告)日：2019-06-04

申请号：US14104139

申请日：2013-12-12

Applicant: AJINOMOTO CO., INC.

Inventor： Daisuke Ejima ,  Haruna Sato ,  Kouhei Tsumoto ,  Masayo Date

IPC: C12P21/00 ,  C07K14/36 ,  C07K16/40 ,  C07K1/113 ,  C12P21/02

Abstract: 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.

公开(公告)号：US20140099672A1

公开(公告)日：2014-04-10

申请号：US14104139

申请日：2013-12-12

Applicant: Ajinomoto Co., Inc.

Inventor： Daisuke Ejima ,  Haruna Sato ,  Kouhei Tsumoto ,  Masayo Date

IPC: C12P21/00

CPC classification number: C12P21/00 ,  C07K1/1136 ,  C07K14/36 ,  C07K16/40 ,  C07K2317/622 ,  C07K2319/00 ,  C12P21/02

Abstract: 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.

Abstract translation: 本发明提供一种由单体蛋白质组成的多聚体蛋白质的制造方法，其特征在于，通过将具有免疫球蛋白折叠结构的蛋白质与能够作为亚单位结构的蛋白质融合而得到的单体蛋白质，其特征在于，包括以下步骤： （A）在微生物的细胞中制备具有不溶性颗粒形式的单体蛋白质; （B）用含有月桂酰-L-Glu的水溶液使步骤（A）中制备的单体蛋白质溶解; （C）在含有精氨酸盐酸盐的缓冲液中稀释步骤（B）中得到的溶液以降低月桂酰-L-Glu的浓度; 和（D）使用凝胶过滤色谱法等用缓冲液代替步骤（C）中得到的溶液的溶剂。