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    IN VITRO PREDICTION OF IN VIVO HALF-LIFE
    1.
    发明申请
    IN VITRO PREDICTION OF IN VIVO HALF-LIFE 审中-公开

    公开(公告)号：US20170227547A1

    公开(公告)日：2017-08-10

    申请号：US15271091

    申请日：2016-09-20

    申请人： Hoffmann-La Roche Inc.

    发明人： Thomas Emrich ,  Hubert Kettenberger ,  Tilman Schlothauer ,  Angela Schoch

    IPC分类号： G01N33/68 ,  B01D15/38 ,  B01D15/16

    CPC分类号： G01N33/6854 ,  B01D15/168 ,  B01D15/3809 ,  C07K16/00 ,  C07K2317/52 ,  C07K2317/94

    摘要： Herein is reported a method for determining the presence of antibody-Fab-FcRn interaction in an antibody-Fc-FcRn complex influencing the in vivo half-life comprising the steps of a) determining the retention time of the antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a first sodium chloride concentration, and b) determining the retention time of the antibody on an FcRn affinity chromatography column with a positive linear pH gradient elution in the presence of a second sodium chloride concentration, whereby the presence of antibody-Fab-FcRn interaction in an antibody-Fc-FcRn complex influencing the in vivo half-life is determined if the retention time determined in step a) and the retention time determined in step b) are substantially different.

    AFFINITY CHROMATOGRAPHY MATRIX
    3.
    发明申请
    AFFINITY CHROMATOGRAPHY MATRIX 有权
    亲属色谱矩阵

    公开(公告)号：US20130338339A1

    公开(公告)日：2013-12-19

    申请号：US13995977

    申请日：2011-12-19

    申请人： Tomas Bjorkman ,  Bjorn Noren ,  Gustav Rodrigo ,  Jelena Vasic ,  Per-Mikael Aberg

    发明人： Tomas Bjorkman ,  Bjorn Noren ,  Gustav Rodrigo ,  Jelena Vasic ,  Per-Mikael Aberg

    IPC分类号： C07K16/12

    CPC分类号： C07K16/1271 ,  B01D15/168 ,  B01D15/3809 ,  B01J20/286 ,  B01J20/289 ,  B01J20/3212 ,  B01J20/3219 ,  B01J20/3274 ,  C07K1/22 ,  C07K14/31

    摘要： The present invention relates to a method of separating one or more immunoglobulin containing proteins from a liquid. The method includes first contacting the liquid with a separation matrix comprising ligands immobilised to a support; allowing the immunoglobulin containing proteins to adsorb to the matrix by interaction with the ligands; followed by an optional step of washing the matrix containing the immunoglobulin containing proteins adsorbed thereon; and recovering said immunoglobulin containing proteins by contacting the matrix with an eluent which releases the proteins. The method improves upon previous separation methods in that each of the ligands comprises one or more of a protein A domain (E, D, A, B, C), or protein Z, or a functional variant thereof, with at least one of the monomers having a substitution of the Asparagine or Histidine at the position corresponding to H18 of B domain of Protein A or Protein Z, and wherein the ligand provides an increase in elution pH compared to non-substituted ligand.

    摘要翻译： 本发明涉及从液体中分离一种或多种含免疫球蛋白的蛋白质的方法。 该方法包括首先使液体与包含固定在载体上的配体的分离基质接触; 允许含有免疫球蛋白的蛋白质通过与配体的相互作用吸附到基质上; 随后是洗涤含有其上吸附有蛋白质的蛋白质的基质的任选步骤; 并通过使基质与释放蛋白质的洗脱液接触来回收所述含有免疫球蛋白的蛋白质。 该方法改进了以前的分离方法，因为每种配体包含蛋白A结构域（E，D，A，B，C）或蛋白质Z或其功能变体中的一种或多种，​​其中至少一种 在相应于蛋白A或蛋白Z的B结构域的H18的位置处取代天门冬酰胺或组氨酸的单体，其中与未取代的配体相比，配体提供了洗脱pH的增加。

    External gradient chromatofocusing
    4.
    发明授权
    External gradient chromatofocusing 失效
    外部梯度色谱聚焦

    公开(公告)号：US07790025B2

    公开(公告)日：2010-09-07

    申请号：US10556562

    申请日：2004-05-14

    申请人： Latchezar I. Tsonev ,  Allen G. Hirsh

    发明人： Latchezar I. Tsonev ,  Allen G. Hirsh

    IPC分类号： B01D15/08

    CPC分类号： B01D15/168 ,  B01D15/362 ,  B01D15/363 ,  B01D15/426 ,  C07K1/16 ,  C07K1/18 ,  C07K1/26 ,  C07K1/28 ,  G01N30/02

    摘要： The invention provides novel methods for the separation of charged molecules such as proteins according to the isoelectric points (pI's) and includes the systems and buffering compositions employed for isolating charged molecules. The invention further provides for modifications to the above described chromatographic methods that enable the separation of charged molecules exhibiting virtually identical pI's by shifting both the buffer's pKa and the pI's of the eluted charged molecules while they are traversing the ion exchange column.

    摘要翻译： 本发明提供了用于根据等电点（pI）分离带电分子如蛋白质的新方法，并且包括用于分离带电分子的系统和缓冲组合物。 本发明进一步提供对上述色谱方法的修改，其能够通过在洗脱的带电荷分子横过离子交换柱的同时移动缓冲液的pKa和洗脱的带电分子的pI来分离表现出实质上相同的pI的带电分子。

    BUFFER KIT AND METHOD OF GENERATING A LINEAR pH GRADIENT
    6.
    发明申请
    BUFFER KIT AND METHOD OF GENERATING A LINEAR pH GRADIENT 有权
    缓冲袋和产生线性pH梯度的方法

    公开(公告)号：US20140179008A1

    公开(公告)日：2014-06-26

    申请号：US13724873

    申请日：2012-12-21

    申请人： Shanhua LIN ,  Christopher A. POHL

    发明人： Shanhua LIN ,  Christopher A. POHL

    IPC分类号： C09K15/30

    CPC分类号： C09K15/30 ,  B01D15/168 ,  B01D15/362 ,  G01N30/34 ,  Y10T436/10 ,  Y10T436/108331 ,  Y10T436/173845 ,  Y10T436/18

    摘要： A buffer kit includes a first eluent and second eluent. The first eluent solution includes at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge, and include a sulfonate group and an amine. The second eluent solution includes at least four buffer salts where at least three of the four buffer salts are a monovalent buffer salt, have a net negative charge or a net neutral zwitterionic charge, and include a sulfonate group and an amine. The first eluent solution has a first pH and the second eluent solution has a second pH where the first pH and second pH are different values. The buffer kit provides a linear pH gradient that forms an approximately straight line from at least the first pH to the second pH.

    摘要翻译： 缓冲试剂盒包括第一洗脱液和第二洗脱液。 第一洗脱溶液包括至少四种缓冲盐，其中四种缓冲盐中的至少三个是一价缓冲盐，具有净负电荷或净中性两性离子电荷，并且包括磺酸盐基和胺。 第二洗脱溶液包括至少四种缓冲盐，其中四种缓冲盐中的至少三个是单价缓冲盐，具有净负电荷或净中性两性离子电荷，并且包括磺酸盐基和胺。 第一洗脱溶液具有第一pH，第二洗脱液具有第二pH，其中第一pH和第二pH是不同的值。 缓冲试剂盒提供线性pH梯度，其从至少第一pH至第二pH形成近似的直线。

    USE OF MIXED MODE CHROMATOGRAPHY FOR THE CAPTURE AND PURIFICATION OF BASIC ANTIBODY PRODUCTS
    8.
    发明申请
    USE OF MIXED MODE CHROMATOGRAPHY FOR THE CAPTURE AND PURIFICATION OF BASIC ANTIBODY PRODUCTS 审中-公开
    混合模式色谱法用于检测和纯化基础抗体产品

    公开(公告)号：US20120208986A1

    公开(公告)日：2012-08-16

    申请号：US13502930

    申请日：2010-10-13

    申请人： Marc D. Wenger ,  Matthew Woodling

    发明人： Marc D. Wenger ,  Matthew Woodling

    IPC分类号： C07K1/16 ,  C07K16/12

    CPC分类号： B01D15/327 ,  B01D15/168 ,  B01D15/362 ,  B01D15/3847 ,  C07K16/065

    摘要： Use of mixed mode chromatography for purification of an antibody from an antibody mixture, for example) a Pichia pastoris fermentation mixture containing impurities such as host cell proteins and DNA is described Mixed mode chromatography is used instead of protein A chromatography and presents certain advantages over protein A chromatography Furthermore, the integration of such a method into a multi-step procedure with other fractionation methods for purification of antibodies suitable for in vivo applications is provided.

    摘要翻译： 使用混合模式色谱法从抗体混合物中纯化抗体，例如）含有诸如宿主细胞蛋白质和DNA的杂质的巴斯德毕赤酵母发酵混合物被描述混合模式色谱用于代替蛋白质A色谱，并且比蛋白质具有某些优点 色谱法此外，还提供了将这种方法整合到具有用于纯化适合于体内应用的抗体的其它分级方法的多步骤方法中。

    Polishing steps used in multi-step protein purification processes
    9.
    发明申请
    Polishing steps used in multi-step protein purification processes 有权
    在多步蛋白纯化过程中使用的抛光步骤

    公开(公告)号：US20070167612A1

    公开(公告)日：2007-07-19

    申请号：US11635800

    申请日：2006-12-06

    申请人： Joe Hua Zhou

    发明人： Joe Hua Zhou

    IPC分类号： C07K16/00

    CPC分类号： C07K16/065 ,  B01D15/168 ,  B01D15/362 ,  B01D15/3809 ,  C07K1/36 ,  G01N30/461 ,  B01D15/363

    摘要： Various embodiments of the present invention are directed to multi-step systems and methods for target-molecule purification that employ column-chromatography-based and/or membrane-filtration-based polishing steps. In one described embodiment of the present invention, a target-protein-containing eluate having a high residual salt concentration is collected from a first chromatography column prepared with an affinity-chromatography resin, loaded onto a second chromatography column prepared with a cation-exchange resin, and eluted from the second cation-exchange column using a buffer in which a time-dependent pH gradient is established. In another described embodiment of the present invention, a partially purified target-protein-containing eluate is collected from a chromatography column and further purified by passing the target-protein-containing eluate through a salt-tolerant anion exchanger.

    摘要翻译： 本发明的各种实施方案涉及采用基于柱层析和/或膜过滤的抛光步骤的靶分子纯化的多步骤系统和方法。 在本发明的一个描述的实施方案中，从用亲和层析树脂制备的第一层析柱收集具有高残留盐浓度的含靶蛋白的洗脱液，将其装载到用阳离子交换树脂制备的第二色谱柱上 ，并使用其中建立时间依赖性pH梯度的缓冲液从第二阳离子交换柱洗脱。 在本发明的另一个描述的实施方案中，从色谱柱收集部分纯化的含靶蛋白的洗脱液，并通过含有靶蛋白的洗脱液通过耐盐阴离子交换剂进一步纯化。