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    METHOD FOR THE PRODUCTION OF DOMAIN ANTIBODIES
    2.
    发明申请
    METHOD FOR THE PRODUCTION OF DOMAIN ANTIBODIES 有权
    产生抗体的方法

    公开(公告)号：US20120157664A1

    公开(公告)日：2012-06-21

    申请号：US13266503

    申请日：2010-04-30

    申请人： Peter Schotte ,  Patrick Stanssens ,  Christine Labeur ,  Jean-Luc Jonniaux ,  Marc Jozef Lauwereys

    发明人： Peter Schotte ,  Patrick Stanssens ,  Christine Labeur ,  Jean-Luc Jonniaux ,  Marc Jozef Lauwereys

    IPC分类号： C12P21/02 ,  C07K16/00

    CPC分类号： C12P21/005 ,  C07K16/00 ,  C07K16/18 ,  C07K16/2875 ,  C07K2317/14 ,  C07K2317/24 ,  C07K2317/35 ,  C07K2317/56 ,  C07K2317/569 ,  C07K2317/624 ,  C07K2317/626

    摘要： The present disclosure relates to a method for producing a domain antibody in a host other than E. coli, preferably yeast, comprising a) applying conditions that promote the formation of disulfide bridges in domain antibodies, or b) removing domain antibodies lacking at least one disulfide bridge, or c) a combination of (a) and (b). More specifically, the present disclosure relates to a method, wherein said conditions that promote the formation of disulfide brides are selected from one or more of the following: a) addition of oxidizing agents, preferably oxidizing metal ions, preferably one or more selected from Cu2+, Fe2+, Fe3+ and Zn2+; b) co-expression of the domain antibody with a thiol isomerase, —c) adapting the culturing conditions by one or more selected from the following: lowering culturing temperature and/or optimizing the culturing medium, including but not limited to reduction of methanol feed for hosts requiring a methanol feed, lowering conductivity of the culture medium, addition of yeast extract and/or peptone, or any combination thereof; d) refolding the domain antibody in the presence of denaturant and redox-buffer e) treating the domain antibody by oxygenation, increasing temperature, increasing pH, high pressure or any combination thereof, and f) combinations of any of a) through e), or wherein conditions that remove domain antibodies lacking at least one disulfide bridge are selected from a) binding domain antibodies comprising free thiol groups to suitable reactive groups, including but not limited to immobilized thiol groups, optionally under denaturing conditions; b) reverse phase high performance chromatography.

    摘要翻译： 本公开内容涉及在除大肠杆菌以外，优选酵母中的宿主中产生结构域抗体的方法，其包括a）施加促进结构域抗体中二硫键形成的条件，或b）除去缺少至少一个 二硫键，或c）（a）和（b）的组合。 更具体地，本公开涉及一种方法，其中促进形成二硫化物新娘的所述条件选自以下一种或多种：a）加入氧化剂，优选氧化金属离子，优选选自Cu 2+的一种或多种， ，Fe2 +，Fe3 +和Zn2 +; b）结构域抗体与硫醇异构酶共表达，-c）使培养条件适应一种或多种选自以下的培养条件：降低培养温度和/或优化培养基，包括但不限于还原甲醇进料 对于需要甲醇进料的主体，降低培养基的导电性，添加酵母提取物和/或蛋白胨或其任何组合; d）在变性剂和氧化还原缓冲液存在下重新折叠结构域抗体e）通过氧合，升高温度，增加pH值，高压或其任何组合来处理结构域抗体，以及f）a）至e）， 或者其中除去缺少至少一个二硫键的结构域抗体的条件选自a）包含游离硫醇基团的结合域抗体适合于适当的反应性基团，包括但不限于固定硫醇基团，任选地在变性条件下; b）反相高效液相色谱法。

    METHODS TO STABILIZE PROTEINS AND POLYPEPTIDES
    3.
    发明申请
    METHODS TO STABILIZE PROTEINS AND POLYPEPTIDES 审中-公开
    稳定蛋白质和多肽的方法

    公开(公告)号：US20110183861A1

    公开(公告)日：2011-07-28

    申请号：US12864969

    申请日：2009-01-29

    申请人： Jean-Luc Jonniaux ,  Marc Jozef Lauwereys ,  Patrick Stanssens

    发明人： Jean-Luc Jonniaux ,  Marc Jozef Lauwereys ,  Patrick Stanssens

    IPC分类号： C40B30/04 ,  C12P21/00 ,  C40B40/10 ,  C07H21/00 ,  C12N1/00 ,  C40B30/00 ,  C07K14/00

    CPC分类号： A61K39/39591 ,  C07K16/36 ,  C07K2317/22 ,  C07K2317/92

    摘要： Methods of modifying and in particular stabilizing proteins and polypeptides by which a predetermined amino acid is introduced into selected positions of said protein or polypeptide to produce a small group of mutants. The methods are based on the premise that certain amino acids play a crucial role in the stability of proteins or polypeptides. Generated mutants can then be further analysed for stability and/or function, e.g. affinity. Furthermore, appropriate mutants may be combined to result in further optimized proteins or polypeptides. In addition, stabilized example polypeptides and suitable methods to identify and/or analyse de-stabilized or stabilized proteins or polypeptides are provided. The methods can be used to study the role of specific amino acids in protein structure and function and to develop new or improved, e.g. stabilized proteins and polypeptides such as antibodies and single variable domains.

    摘要翻译： 修饰和特别是稳定蛋白质和多肽的方法，通过该蛋白质和多肽将预定的氨基酸引入所述蛋白质或多肽的选定位置以产生少量的突变体。 这些方法基于某些氨基酸在蛋白质或多肽的稳定性中发挥关键作用的前提。 然后可以进一步分析产生的突变体的稳定性和/或功能，例如。 亲和力。 此外，合适的突变体可以组合以产生进一步优化的蛋白质或多肽。 此外，提供了稳定的实施例多肽和鉴定和/或分析去稳定化或稳定的蛋白质或多肽的合适方法。 该方法可用于研究特定氨基酸在蛋白质结构和功能中的作用，并开发新的或改进的，例如， 稳定的蛋白质和多肽，例如抗体和单可变结构域。

    STABLE FORMULATIONS OF POLYPEPTIDES AND USES THEREOF
    7.
    发明申请
    STABLE FORMULATIONS OF POLYPEPTIDES AND USES THEREOF 有权
    稳定的多肽制剂及其用途

    公开(公告)号：US20120201812A1

    公开(公告)日：2012-08-09

    申请号：US13393679

    申请日：2010-09-03

    申请人： Ann Brige ,  Christine Labeur ,  Marc Jozef Lauwereys

    发明人： Ann Brige ,  Christine Labeur ,  Marc Jozef Lauwereys

    IPC分类号： A61K39/395 ,  A61P19/08 ,  A61P19/10

    CPC分类号： A61K39/39591 ,  A61K9/0019 ,  A61K47/02 ,  A61K47/14 ,  A61K47/26

    摘要： Stable formulations are provided that contain immunoglobulin single variable domains at a high concentration. The formulations are useful as pharmaceutical formulation and suitable for subcutaneous administration. The formulations can be transported and stored under various stress conditions. The invention further relates to containers and pharmaceutical units comprising such formulations and to methods for preparing and prophylactic and therapeutic uses of the formulations and pharmaceutical units of the invention.

    摘要翻译： 提供了稳定的制剂，其含有高浓度的免疫球蛋白单可变结构域。 该制剂可用作药物制剂并且适于皮下给药。 制剂可以在各种压力条件下运输和储存。 本发明还涉及包含这些制剂的容器和药物单元以及本发明的制剂和药物单元的制备和预防和治疗用途的方法。