Immunoglobulin-binding protein and affinity carrier using same

A technology of immunoglobulin and protein, which is applied in the field of immunoglobulin binding protein, can solve the problems such as improving the binding capacity of immunoglobulin of the carrier, and achieve dynamic binding capacity that is not easy to decrease, high and low binding capacity of immunoglobulin cost effect

Pending Publication Date: 2017-12-01
JSR CORPORATIOON +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned prior art is mainly aimed at improving the alkali resistance of the carrier, and does not mention improving the immunoglobulin binding ability of the carrier.

Method used

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  • Immunoglobulin-binding protein and affinity carrier using same
  • Immunoglobulin-binding protein and affinity carrier using same
  • Immunoglobulin-binding protein and affinity carrier using same

Examples

Experimental program
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Embodiment

[0101] Hereinafter, the present invention will be described in more detail with reference to examples. In addition, the following description generally shows the form of this invention, and unless otherwise indicated, this invention is not limited to this description.

reference example 1

[0102] Reference Example 1 Synthesis of Porous Particles

[0103] Dissolve 8.2 g of glycidyl methacrylate (manufactured by MITSUBISHI RAYON), 65.9 g of trimethylolpropane trimethacrylate (manufactured by Sartomer), and 90.6 g of glycerin monomethacrylate (manufactured by NOF) in To 245.8 g of 2-octanone (manufactured by Toyosei Kogyo Co., Ltd.) and 62 g of acetophenone (manufactured by Wako Pure Chemical Industries, Ltd.), 2 g of 2,2'-azobisisobutyronitrile (manufactured by Wako Pure Chemical Industries, Ltd.) was added, Prepare the organic monomer solution.

[0104] Next, 8.5 g of polyvinyl alcohol (PVA-217 manufactured by Kuraray Co., Ltd.), 0.43 g of sodium lauryl sulfate (Emal 10G manufactured by Kao Corporation), and sodium sulfate (manufactured by Wako Pure Chemical Industries, Ltd.) were added to 4240 g of pure water. 21.3 g, stirred overnight to prepare an aqueous solution.

[0105] Next, the obtained aqueous solution was poured into a 7L separable flask, a thermomet...

Embodiment 1

[0107] Example 1 Production of C domain recombinant immunoglobulin binding protein 1 (IgGBPC1)

[0108] A plasmid encoding the amino acid sequence shown in SEQ ID NO: 4 was prepared, and Escherichia coli competent cells BL21(DE3) (manufactured by STRATAGENE) were transformed using this plasmid to obtain a recombinant. The resulting recombinant was incubated at 37°C until the absorbance (OD600) reached about 10. Thereafter, IPTG (manufactured by Sigma-Aldrich) was added to a final concentration of 1 mM, and further incubated at 37° C. for 4 hours to express the recombinant immunoglobulin-binding protein. After protein expression, cells were recovered and disrupted in Tris buffer, pH 9.5. Immunoglobulin-binding proteins were purified from the obtained cell lysates by anion-exchange chromatography (Q-Sepharose FF, manufactured by GE Health Care Bio-Sciences) and cation-exchange chromatography (SP-Sepharose FF, manufactured by GE Health Care Bio-Sciences). Purified immunoglobuli...

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Abstract

Provided is an affinity chromatography carrier in which high immunoglobulin-binding capacity and alkali resistance are maintained. Provided is an immunoglobulin-binding protein, wherein: the immunoglobulin-binding protein contains at least one modified immunoglobulin-binding domain; and the modified immunoglobulin-binding domain is a polypeptide comprising an amino acid sequence having at least one amino acid residue inserted between positions corresponding to 3 and 4 of the amino acid sequence of the B domain, Z domain, or C domain in an amino acid sequence of an immunoglobulin-binding domain selected from the group consisting of the B domain, Z domain, C domain, and variants thereof of Staphylococcus aureus protein A.

Description

technical field [0001] The present invention relates to an immunoglobulin-binding protein, an affinity carrier using the same, and a method for isolating an immunoglobulin and a method for producing an antibody drug using the affinity carrier. Background technique [0002] Affinity chromatography is a chromatography method using a column filled with a ligand-immobilized carrier in which a substance (ligand) that specifically binds to a substance for separation or purification is immobilized on an insoluble carrier And get. Affinity chromatography is used for separation and purification of biologically related substances such as proteins and nucleic acids (Patent Document 1). As the support for affinity chromatography, for example, cross-linked particles of sugar chains represented by agarose gel and particles mainly composed of synthetic polymers have been used. [0003] In preparation of a support for affinity chromatography, it is necessary to immobilize a ligand, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C07K1/22C07K14/195C07K17/04C12N1/15C12N1/19C12N1/21C12N5/10C12P21/02
CPCC07K1/22C07K17/04C12N5/10C12N15/09C07K14/31G01N33/6854C07K16/065C07K14/195C07K16/1203C07K17/08
Inventor 安冈润一市居敬中村聪塩谷知范板谷佳织
Owner JSR CORPORATIOON
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