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    METHOD FOR THE PURIFICATION OF AT LEAST ONE TARGET SUBSTANCE THAT IS TO BE IDENTIFIED
    1.
    发明申请
    METHOD FOR THE PURIFICATION OF AT LEAST ONE TARGET SUBSTANCE THAT IS TO BE IDENTIFIED 审中-公开
    法提纯至少一个目标物质

    公开(公告)号：WO2008055671A3

    公开(公告)日：2008-09-18

    申请号：PCT/EP2007009648

    申请日：2007-11-07

    Applicant: PETER JOCHEN

    Inventor： PETER JOCHEN

    IPC: G01N33/543 ,  C12N5/00

    CPC classification number: G01N33/54333 ,  C12N5/0018

    Abstract: Disclosed is a method for purifying at least one target substance that is to be identified and is present or is formed in a cell culture medium when cells are cultivated. In said method, magnetic particles, i.e. beads, to the functionalized surface of which the target substance selectively attaches, are added to the cell culture medium, and the particles to which the target substance is attached are selected out of the cell culture medium by applying a magnetic field. The method is characterized by the following steps: a serum substitute is provided that is obtained from a natural serum and is free or virtually free of low-molecular substances having a maximum mass of 60 kDa, particularly a maximum mass of 10 kDa; the serum substitute is admixed to the cell culture medium which already contains the cells or to which the cells are added; the cells are incubated in the cell culture medium enriched with serum substitute; at least some of the cell culture supernatant formed during the incubation is separated; the cell culture supernatant is filtered by means of an ultrafiltering process so as to obtain a retentate; the beads are supplied in such a way that the functionalized surface of the beads comprises a plurality of dendrimers containing up to 10 branches, i.e. ten generations, the terminal points of the last generation of each dendrimer being modified; the beads and the retentate are admixed to a buffer solution so as to obtain a mixture; the target substances contained in the retentate are incubated and are fixed to the beads; and the magnetic beads are magnetically selected out of the mixture.

    Abstract translation: 用于纯化的方法被描述的目标物质，所述während.einer细胞培养在细胞培养介质中的至少发生或形成，其中细胞培养基中的磁性官能在其表面上的粒子，即所谓的小珠，被添加到其表面的目标物质选择性地 退火并通过施加磁场，有损粒子与目标物质从细胞培养基中进行选择。 该方法的特征在于以下步骤：提供由天然存在的血清回收的，并且不含或几乎不含是低分子量物质，其具有小于或等于60千道尔顿，特别是小于或血清替代物的等于10kDa的质量，具有，混合所述 血清替代品，其中所述细胞包含或加入到其中的细胞中，细胞培养基中温育细胞的含血清替代细胞培养基富集的，分离至少一部分自调节温育细胞培养物上清液中，通过超滤获得过滤细胞培养物上清液 滞留物提供所述珠，使得多个树枝状聚合物的含多至10个分支的小珠，即，官能化表面 十代，其中所述最后一代树枝状聚合物的末端点被修饰的，混合的珠和在缓冲液中渗余物以形成混合物，温育并结合靶物质包含在渗余物到珠子和从磁珠磁性选择 混合物。

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