Method of enhancing the performance of chromatography methods for purification of proteins

    公开(公告)号:US10927145B2

    公开(公告)日:2021-02-23

    申请号:US16309889

    申请日:2017-06-12

    Abstract: This application describes compositions, kits, and methods for removing contaminants from a protein preparation, for example a cell culture harvest comprising a recombinant protein. The methods described herein can enhance the performance of a later applied chromatographic purification step. The method generally involves contacting a protein preparation comprising a desired protein with a combination of allantoin, a cationic polymer, and a fatty acid, thereby causing the formation of solids and removal of the solids using a suitable process. Further treatment by contacting the treated sample with a chemically reactive surface further enhances results achieved by the first purification step.

    Protein purification methods
    4.
    发明授权

    公开(公告)号:US09988419B2

    公开(公告)日:2018-06-05

    申请号:US14766131

    申请日:2014-02-05

    Abstract: A method for the purification of a desired protein from a protein preparation includes conditioning the protein preparation by treatment with soluble organic multivalent ions, immobilized organic multivalent ions, or both, optionally in the presence of supersaturated allantoin, thereby removing at least 90% of chromatin, then (1) precipitating the desired protein with a nonionic organic polymer in the presence of non-protein-precipitating salts at greater than physiological concentration to provide a precipitate of the desired protein; or (2) precipitating the desired protein with a nonionic organic polymer in the absence of non-precipitating salts at greater than physiological concentration to provide a precipitate and subsequently washing the precipitate with a nonionic organic polymer in the presence of non-protein-precipitating salts at greater than physiological concentration.

    CHROMATOGRAPHIC PURIFICATION OF IMMUNOGLOBULIN G PREPARATIONS WITH PARTICLES HAVING MULTIMODAL FUNCTIONALITIES
    8.
    发明申请
    CHROMATOGRAPHIC PURIFICATION OF IMMUNOGLOBULIN G PREPARATIONS WITH PARTICLES HAVING MULTIMODAL FUNCTIONALITIES 有权
    具有多种功能的颗粒的免疫球蛋白G制备的色谱纯化

    公开(公告)号:US20150183852A1

    公开(公告)日:2015-07-02

    申请号:US14555060

    申请日:2014-11-26

    Abstract: A method of purifying a sample containing a desired protein includes the steps of (i) providing a packed chromatographic column having positively charged porous particles, (ii) equilibrating the column to the conditions to which the desired protein in the sample is to elute, (iii) contacting the sample with the packed chromatographic column such that the sample volume applied to the packed chromatographic column is less than or equal to the interparticle space of the positively charged porous particles within the packed chromatographic column, (iv) eluting the desired protein from the packed chromatographic column, where the desired protein is in a purer state and in the conditions to which the packed chromatographic column was equilibrated; where the desired protein is an antibody, an antibody fragment, an antibody derivative, or an antibody fusion protein.

    Abstract translation: 纯化含有所需蛋白质的样品的方法包括以下步骤:(i)提供具有带正电荷的多孔颗粒的填充色谱柱,(ii)将色谱柱平衡至样品中所需蛋白质洗脱的条件( iii)使样品与填充色谱柱接触,使得施加到填充色谱柱的样品体积小于或等于填充色谱柱中带正电荷的多孔颗粒的颗粒间间隔,(iv)从所需的蛋白质洗脱所需的蛋白质 填充色谱柱,其中所需的蛋白质处于更纯的状态和填充色谱柱平衡的条件下; 其中所需蛋白质是抗体,抗体片段,抗体衍生物或抗体融合蛋白。

    Protein purification in the presence of nonionic organic polymers and electropositive surfaces

    公开(公告)号:US10112971B2

    公开(公告)日:2018-10-30

    申请号:US14769101

    申请日:2014-02-26

    Abstract: A method of purifying a desired protein from a preparation includes (a) providing the preparation in a form having less than about 5% of chromatin residing in an original production medium, (b) contacting the preparation with a nonionic organic polymer and a salt, a concentration of nonionic organic polymer being sufficient to precipitate the desired protein or cause its accretion on a hydrophilic surface, or maintain it in a precipitated state or accreted on the hydrophilic surface, the salt concentration being sufficient to produce greater than physiological conductivity, and (c) contacting the preparation with at least one electropositive surface, optionally in the presence of a salt concentration sufficient to produce greater than physiological conductivity, the desired protein does not substantially adsorb to the at least one electropositive surface while not preventing adsorption of acidic contaminants to the at least one electropositive surface.

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