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Purification method for divalent cation binding proteins on anion exchange resin

A divalent cation and protein-binding technology, applied in the field of purification of divalent cation-binding proteins on anion exchange resins

Inactive Publication Date: 2013-04-03
BAXALTA INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Despite the successful isolation of bovine factor VII from bovine plasma, the purification of human factor VII is problematic, i.e., the material produced is only partially pure or is obtained in such small quantities that it can be characterized as active without detectable protein

Method used

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  • Purification method for divalent cation binding proteins on anion exchange resin
  • Purification method for divalent cation binding proteins on anion exchange resin
  • Purification method for divalent cation binding proteins on anion exchange resin

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Embodiment Construction

[0013] In one aspect, the invention relates to a method of purifying a divalent cation binding protein comprising the steps of:

[0014] (a) Loading the divalent cation-binding protein to the anion exchange resin material in a loading buffer in the absence of divalent cations, optionally washing the loaded anion exchange resin material with a wash buffer in the absence of divalent cations ;and

[0015] (b) eluting the divalent cation-binding protein with an eluent comprising at least one divalent cation to form an eluate comprising the divalent cation-binding protein;

[0016] Wherein the pH of the eluent in step (b) is higher than the pH of the washing buffer in step (a), or in the absence of a washing step, higher than the pH of the loading buffer in step (a).

[0017] As used herein, the term "absence of divalent cations" refers to the absence of free divalent cations and any protein-bound divalent cations in the buffer, where divalent cations complexed with or by a chelat...

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Abstract

The present invention relates to a method for the purification of divalent cation binding proteins with high purity on an anion exchange resin material, to divalent cation binding proteins obtainable by said method, and to a kit comprising means for carrying out said method.

Description

field of invention [0001] The present invention relates to a method for the purification of divalent cation-binding proteins to high purity on anion-exchange resin materials, to divalent cation-binding proteins obtainable by said method and to a kit comprising means for carrying out said method. Background of the invention [0002] To date, many mammalian proteins are produced in host cells by, for example, transfecting the cells with DNA encoding the protein and growing the recombinant cells under conditions favorable for expression of the protein. Proteins secreted by cells into the cell culture medium or present within the cells can be separated from the medium and other components using chromatographic techniques, eg, ion exchange chromatography, affinity chromatography, and the like. Purification is especially important for further pharmaceutical applications. However, after the target protein is completely purified, the biological activity of the protein must be maint...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/18C12N9/64
CPCC12N9/647C07K14/4721C12N9/6437C12Y304/21021C12Y304/21022C07K1/18C12N9/644A61P7/04C07K1/14C07K14/47C12N9/64
Inventor A.米特雷尔M.哈斯拉切尔C.菲德勒
Owner BAXALTA INC
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