AFFINITY-ASSISTED PROTEIN MODIFICATION AND RECYCLING

摘要

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.