[EN] AFFINITY-ASSISTED PROTEIN MODIFICATION AND RECYCLING<br/>[FR] MODIFICATION ET RECYCLAGE DE PROTÉINES BASÉS SUR LEUR AFFINITÉ
申请人:UNIV CALIFORNIA
公开号:WO2016118770A1
公开(公告)日:2016-07-28
The present invention provides methods for preparing a protein conjugate having a defined number of conjugate groups. The method includes: forming a mixture containing a macrocyclic matrix material and a plurality of proteins; eluting the proteins from the macrocyclic matrix material to obtain a first separated protein fraction and a second separated protein fraction, wherein substantially all of the proteins in the first separated protein fraction have the same number of handle moieties; contacting the handle moieties with a conversion reagent under conditions sufficient to convert the handle moieties in the first separated protein fraction to reactive moieties; and contacting the reactive moieties with a conjugation reagent under conditions sufficient to form a plurality of protein conjugates, wherein substantially all of the protein conjugates in the plurality have the same number of conjugate groups. Also disclosed are methods for recovering enzymes and other proteins from mixtures for isolation and/or reuse of the enzymes and proteins.
本发明提供了制备具有定义数量结合基团的蛋白共轭物的方法。该方法包括:形成含有大环矩阵材料和多种蛋白质的混合物;从大环矩阵材料中洗脱蛋白质,以获得第一分离的蛋白质分数和第二分离的蛋白质分数,其中第一分离的蛋白质分数中几乎所有蛋白质具有相同数量的手柄基团;将手柄基团与转化试剂接触,在足以将第一分离的蛋白质分数中的手柄基团转化为反应性基团的条件下;将反应性基团与结合试剂接触,在足以形成多种蛋白共轭物的条件下,其中多数蛋白共轭物具有相同数量的结合基团。还公开了从混合物中回收酶和其他蛋白质以便分离和/或重复使用酶和蛋白质的方法。