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    • 51. 发明申请
    • NEW CHELATORS FOR AFFINITY PURIFICATION OF RECOMBINANT PROTEINS
    • 新型重组蛋白的亲和性纯化剂
    • WO2017046625A1
    • 2017-03-23
    • PCT/IB2015/002003
    • 2015-06-25
    • CUBE BIOTECH GMBH
    • B01J45/00B01J20/289B01J20/32
    • B01J20/289B01J20/3204B01J20/3212B01J20/3219B01J20/3265B01J45/00
    • The present invention relates to a method of preparing a solid-phase bound multimeric chelator where the chelator is coupled via one carboxylic group. The chelator consists of aminopolycarboxylic acid, connected to other aminocarboxylic acids via di-or trifunctional linkers. Examples for this invention have the formulas (I): Wherein Z is a solid phase, S is an optional spacer molecule, able to bind both to the solid phase and the chelator, APA1 is a tetradentate aminopolycarboxylic acid, K is a bifunctional linker molecule, L is a trifunctional linker molecule, APA2 is a pentadentate aminopolycarboxylic acid, n is 1 - 50. Furthermore, the invention relates to a method for purifying his-tagged recombinant polypeptides and proteins, phosphoproteins and metal-coordinating proteins with solid phase-bound di-, tri- and tetra- and polymeric chelators. These chelators are polycarboxylates, such as EDTA, DTPA, and the like, and they are connected via linkers, forming chains. Coupling groups to the linker are selected from ester-, thioester- or amide functionalities. The chelator chains are bound to a solid phase via an ester-, thioester- or amide group of one terminal chelator. The solid-phase bound chelator chains, loaded with metal ions, provide a strong metal binding, high stability against chelators and reductants, high affinity and an extremely high binding capacity.
    • 本发明涉及一种制备固相结合的多聚体螯合剂的方法,其中螯合剂通过一个羧基连接。 螯合剂由氨基多羧酸组成,通过二 - 或三官能接头与其它氨基羧酸连接。 本发明的实例具有式(I):其中Z是固相,S是能够结合固相和螯合剂的任选的间隔分子,APA1是四齿氨基多羧酸,K是双官能连接分子 ,L是三官能连接分子,APA2是五齿多氨基多羧酸,n为1-50。此外,本发明涉及一种用于纯化其标签的重组多肽和蛋白质,磷蛋白和具有固相结合的金属配位蛋白的方法 二,三和四和聚合螯合剂。 这些螯合剂是多羧酸盐,例如EDTA,DTPA等,它们通过连接体连接形成链。 连接基团与连接体的偶联选自酯 - ,硫酯 - 或酰胺官能团。 螯合剂链通过一个末端螯合剂的酯 - ,硫酯 - 或酰胺基团结合到固相。 负载有金属离子的固相螯合剂链提供强烈的金属结合,对螯合剂和还原剂的高稳定性,高亲和力和极高的结合能力。