公开(公告)号：US09643174B2

公开(公告)日：2017-05-09

申请号：US14542686

申请日：2014-11-17

Applicant: ASAHI KASEI MEDICAL CO., LTD.

Inventor： Ichiro Koguma ,  Hiroki Shigematsu ,  Kazuo Okuyama ,  Teruo Okano ,  Yoshikatsu Akiyama ,  Kenichi Nagase

IPC: B01J39/20 ,  C07K1/18 ,  B01J39/26 ,  C08F293/00 ,  B01J20/285 ,  B01J20/32 ,  B01J39/05 ,  B01J39/19

CPC classification number: B01J39/20 ,  B01J20/285 ,  B01J20/3276 ,  B01J20/3278 ,  B01J39/05 ,  B01J39/19 ,  B01J39/26 ,  C07K1/18 ,  C08F293/005 ,  C08F2438/01

Abstract: Provided is a temperature responsive adsorbent prepared by immobilizing a copolymer containing at least N-isopropylacrylamide to a base material surface. The copolymer has at least a strong cation exchange group. In addition, the copolymer contains the strong cation exchange group in an amount of 0.01 to 5 mol % relative to N-isopropylacrylamide in terms of monomer.

公开(公告)号：US20150239956A1

公开(公告)日：2015-08-27

申请号：US14408458

申请日：2013-06-27

Applicant: ASAHI KASEI MEDICAL CO., LTD. ,  NOMADIC BIOSCIENCE CO., LTD.

Inventor： Ichiro Koguma ,  Kazuo Okuyama ,  Satoshi Sato

IPC: C07K16/00

CPC classification number: C07K16/00 ,  C07K1/22 ,  C07K16/18 ,  C07K16/30 ,  C07K2317/21 ,  C07K2317/92

Abstract: A method for manufacturing a high-affinity antibody including purifying an antibody to which a treatment of reducing the affinity for an antigen is not applied by a temperature responsive protein A medium, in which dissociation constant (KD value) to an antigen is smaller than the KD value of an antibody purified by an acid elution type protein A medium.

Abstract translation: 一种制备高亲和力抗体的方法，包括纯化抗体，其中抗原抗体的亲和力降低的抗体不通过对抗原的解离常数（KD值）小于 通过酸洗脱型蛋白A培养基纯化的抗体的KD值。

公开(公告)号：US20150218208A1

公开(公告)日：2015-08-06

申请号：US14423568

申请日：2013-08-27

Applicant: ASAHI KASEI MEDICAL CO., LTD.

Inventor： Ichiro Koguma ,  Rumiko Ishikawa ,  Hiroshi Takemasa ,  Kazuo Okuyama

IPC: C07K1/22

CPC classification number: C07K1/22 ,  B01D15/3809 ,  B01D15/3876 ,  G01N2030/8813 ,  B01D15/362

Abstract: There is provided a method for purifying an antibody using temperature-responsive Protein A, wherein different buffer solutions are used in a washing step of washing a stationary phase having the temperature-responsive Protein A and in an elution step of eluting the antibody captured by the stationary phase, the method including the washing step of washing the stationary phase using a buffer solution at a temperature at which the temperature-responsive Protein A and the antibody are bound and having a first salt concentration, and the elution step of eluting the antibody captured by the stationary phase using a buffer solution at a temperature at which the antibody is released from the temperature-responsive Protein A and having a second salt concentration lower than the first salt concentration.

Abstract translation: 提供了使用温度敏感性蛋白A纯化抗体的方法，其中在洗涤具有温度敏感性蛋白A的固定相的洗涤步骤中使用不同的缓冲溶液，并且在洗脱步骤中洗脱由 所述方法包括在温度响应性蛋白A和抗体结合并具有第一盐浓度的温度下使用缓冲溶液洗涤固定相的洗涤步骤，以及洗脱被捕获的抗体的洗脱步骤 通过使用缓冲溶液在抗体从温度敏感性蛋白A释放并具有低于第一盐浓度的第二盐浓度的温度下通过固定相。

公开(公告)号：US20150191506A1

公开(公告)日：2015-07-09

申请号：US14410616

申请日：2013-06-28

Applicant: ASAHI KASEI MEDICAL CO., LTD. ,  NOMADIC BIOSCIENCE CO., LTD.

Inventor： Kazuo Okuyama ,  Ichiro Koguma ,  Tomokiyo Marumoto ,  Satoshi Sato

IPC: C07K1/22 ,  B01J20/24 ,  C08F16/06 ,  C07K16/00 ,  C07K14/195

CPC classification number: C07K1/22 ,  B01D15/3809 ,  B01D15/3876 ,  B01J20/24 ,  C07K14/195 ,  C07K16/00 ,  C08F16/06 ,  Y10T428/2982

Abstract: It is an object of the present invention to provide an affinity chromatographic adsorbent using temperature-responsive protein A, wherein the adsorbent is capable of improving the culture productivity of the temperature-responsive protein A and the stability of the temperature-responsive protein A in cell disruption solution. According to the present invention, there is provided an adsorbent consisting of a carrier, to which a polypeptide comprising a tag peptide, a linker sequence, and a B-domain mutant derived from protein A from the N-terminal side thereof binds, wherein the linker sequence is an amino acid sequence that does not comprise a Val-Pro-Arg sequence and is composed of 7 to 12 amino acid residues; and the binding property of the B-domain mutant derived from protein A to an immunoglobulin can vary depending on temperature under conditions of pH 5 to 9 and a temperature of lower than 60° C.

Abstract translation: 本发明的目的是提供使用温度敏感性蛋白A的亲和层析吸附剂，其中吸附剂能够提高温度响应性蛋白A的培养生产率和细胞中温度敏感性蛋白A的稳定性 破解解决方案 根据本发明，提供了一种由载体组成的吸附剂，其包含标签肽的多肽，接头序列和从N-末端侧衍生自蛋白A​​的B结构域突变体结合，其中 接头序列是不包含Val-Pro-Arg序列并由7至12个氨基酸残基组成的氨基酸序列; 并且衍生自蛋白A​​的B结构域突变体与免疫球蛋白的结合性质可以根据pH5-9的条件下的温度和低于60℃的温度而变化。