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    PURIFICATION OF POLYPEPTIDES USING DUAL STAGE TANGENTIAL-FLOW ULTRAFILTRATION
    6.
    发明授权
    PURIFICATION OF POLYPEPTIDES USING DUAL STAGE TANGENTIAL-FLOW ULTRAFILTRATION 有权

    公开(公告)号：EP2914612B1

    公开(公告)日：2018-11-14

    申请号：EP13794828.7

    申请日：2013-10-28

    Applicant: F. Hoffmann-La Roche AG

    Inventor： BECKER, Peter ,  NEUMANN, Sebastian

    IPC: C07K1/34

    CPC classification number: C07K1/34 ,  B01D15/362 ,  B01D15/363 ,  B01D61/145 ,  B01D2315/10 ,  B01D2317/02 ,  C07K1/36 ,  C12M47/10 ,  C12M47/12

    Abstract: The present invention is directed to methods for the separation of a molecule of interest from a solution containing the molecule using dual stage tangential-flow ultrafiltration (“TFF”). In particular, the methods of the invention are directed to the processing of crude feed streams such as conditioned cell culture supernatant to dramatically reduce contaminant and/or impurity levels prior to subsequent, i.e., downstream, refining unit operations. The methods of the invention may be used in the processing of a crude feed stream from biological production systems such as fermentation or other cell culture process, and may further eliminate the need for time consuming impurity precipitation (e.g., pH driven) and/or precipitate filtration processes prior to downstream processes that are sensitive to high impurity loads such as chromatographic unit operations. The disclosed dual stage TFF process combines at least two TFF unit operations that may be advantageously conducted at a pH that corresponds to or is about that of the pH of the feed stream, e.g., a cell culture supernatant, typically a pH of 7.5±1.0. The use of the TFF unit operations to supplement, improve or replace traditional processes for purification of proteins of interest for a feed stream may represent significant savings in both direct and indirect processing costs, For example, in addition to indirect savings by eliminating precipitation and precipitate filtration processes, the reduction in impurity loads effected by the dual stage TFF unit operations may result in indirect savings by improving downstream column performance, e.g., chromatographic separation, dynamic binding capacity, operational lifetime and/or a reduction of the required column size. In particular embodiments, the methods of the invention are used in processes for the purification of immunoglobulin molecules, e.g., antibodies, which processes are devoid of affinity purification steps, e.g., protein A affinity chromatography purification.

    CATION AND ANION EXCHANGE CHROMATOGRAPHY METHOD
    8.
    发明授权
    CATION AND ANION EXCHANGE CHROMATOGRAPHY METHOD 有权

    公开(公告)号：EP2748178B1

    公开(公告)日：2018-10-31

    申请号：EP12748689.2

    申请日：2012-08-17

    Applicant: F. Hoffmann-La Roche AG

    Inventor： FALKENSTEIN, Roberto ,  MAGRI, Maria, Laura ,  MEHR, Michaela ,  SCHWENDNER, Klaus ,  SPENSBERGER, Bernhard

    IPC: C07K1/18

    CPC classification number: C07K14/775 ,  B01D15/362 ,  B01D15/363 ,  C07K1/18

    Abstract: Herein is reported a method for purifying a polypeptide comprising the steps of i) applying a solution comprising the polypeptide to an ion exchange chromatography material, and ii) recovering the polypeptide with a solution comprising a denaturant and thereby purifying the polypeptide, whereby the ion exchange chromatography material comprises a matrix of cross-linked poly (styrene-divinylbenzene) to which ionic ligands have been attached, and wherein the solution comprising the polypeptide applied to the ion exchange chromatography material is free of the denaturant and the polypeptide adsorbed to the ion exchange chromatography material is recovered with a solution comprising a denaturant at a constant conductivity.

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