公开(公告)号：US20150152195A1

公开(公告)日：2015-06-04

申请号：US14406728

申请日：2013-06-11

Applicant: DAISO CO., LTD.

Inventor： Masahiro Iwakura ,  Kiyonori Hirota ,  Seiji Otaka

IPC: C07K17/14 ,  C07K1/22 ,  B01D15/38 ,  C07K14/31

CPC classification number: C07K17/14 ,  B01D15/3809 ,  B01J20/286 ,  B01J20/3204 ,  B01J20/3217 ,  B01J20/3274 ,  C07K1/22 ,  C07K14/31 ,  C07K16/065 ,  C07K2317/92

Abstract: An object of the present invention is to provide a carrier on which Protein A has been immobilized, the Protein A having a specific amino acid sequence that permits the desorption of antibodies under mild pH conditions (specifically, pH4.0 to 5.5) in which many antibodies do not undergo acid modification; and a manufacturing method for the carrier. An immobilization carrier (excluding an immobilization carrier having a monolith structure) on which a protein is adsorbed by electrostatic interaction, the protein consisting of an amino acid sequence represented by the general formula: R1-R2-R3-R4-R5-R6, wherein the part represented by R1-R2-R3 is used for immobilization on the immobilization carrier, wherein: the sequence represents a sequence from the amino-terminal side towards the carboxyl-terminal side; the sequence of the R2 part is the sequence of a Protein A mutant as a protein to be immobilized or a sequence in which 1 to 3 units of the sequence thereof were linked together, the Protein A mutant having characteristics of strongly binding to an antibody under neutral conditions and dissociating with the antibody bound under neutral conditions under weakly acidic conditions of pH 4.0 to 5.5.

Abstract translation: 本发明的目的是提供一种其上固定有蛋白A的载体，具有特定氨基酸序列的蛋白A，其允许在温和的pH条件（具体地，pH4.0至5.5）下解吸抗体，其中许多 抗体不经过酸修饰; 和载体的制造方法。 通过静电相互作用吸附有蛋白质的固定载体（不包括具有整料结构的固定化载体），由通式R1-R2-R3-R4-R5-R6表示的氨基酸序列组成的蛋白质，其中 由R1-R2-R3表示的部分用于固定化载体上，其中：该序列表示从氨基末端侧到羧基末端侧的序列; R2部分的序列是作为要固定的蛋白质的蛋白A突变体的序列或其中1至3个单元的序列连接在一起的序列，具有与抗体强烈结合的特征的蛋白A突变体 中性条件，并在pH 4.0至5.5的弱酸性条件下，在中性条件下与抗体结合解离。

公开(公告)号：US09534060B2

公开(公告)日：2017-01-03

申请号：US14406728

申请日：2013-06-11

Applicant: DAISO CO., LTD.

Inventor： Masahiro Iwakura ,  Kiyonori Hirota ,  Seiji Otaka

IPC: C07K17/14 ,  C07K1/22 ,  C07K14/31 ,  C07K16/06 ,  B01D15/38 ,  B01J20/286 ,  B01J20/32

CPC classification number: C07K17/14 ,  B01D15/3809 ,  B01J20/286 ,  B01J20/3204 ,  B01J20/3217 ,  B01J20/3274 ,  C07K1/22 ,  C07K14/31 ,  C07K16/065 ,  C07K2317/92

Abstract: An object of the present invention is to provide a carrier on which Protein A has been immobilized, the Protein A having a specific amino acid sequence that permits the desorption of antibodies under mild pH conditions (specifically, pH 4.0 to 5.5) in which many antibodies do not undergo acid modification; and a manufacturing method for the carrier. An immobilization carrier (excluding an immobilization carrier having a monolith structure) on which a protein is adsorbed by electrostatic interaction, the protein consisting of an amino acid sequence represented by the general formula: R1-R2-R3-R4-R5-R6, wherein the part represented by R1-R2-R3 is used for immobilization on the immobilization carrier, wherein: the sequence represents a sequence from the amino-terminal side towards the carboxyl-terminal side; the sequence of the R2 part is the sequence of a Protein A mutant as a protein to be immobilized or a sequence in which 1 to 3 units of the sequence thereof were linked together, the Protein A mutant having characteristics of strongly binding to an antibody under neutral conditions and dissociating with the antibody bound under neutral conditions under weakly acidic conditions of pH 4.0 to 5.5.

Abstract translation: 本发明的一个目的是提供一种其上固定有蛋白A的载体，具有特定氨基酸序列的蛋白A，其允许在温和的pH条件（特别是pH 4.0至5.5）下解吸抗体，其中许多抗体 不经酸改性; 和载体的制造方法。 通过静电相互作用吸附有蛋白质的固定载体（不包括具有整料结构的固定化载体），由通式R1-R2-R3-R4-R5-R6表示的氨基酸序列组成的蛋白质，其中 由R1-R2-R3表示的部分用于固定化载体上，其中：该序列表示从氨基末端侧到羧基末端侧的序列; R2部分的序列是作为要固定的蛋白质的蛋白A突变体的序列或其中1至3个单元的序列连接在一起的序列，具有与抗体强烈结合的特征的蛋白A突变体 中性条件，并在pH 4.0至5.5的弱酸性条件下，在中性条件下与抗体结合解离。