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Syntheses and applications of water‐soluble reactive polymers for purification and immobilization of biomolecules
Author(s) -
Nguyen A. L.,
Luong J. H. T.
Publication year - 1989
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260340909
Subject(s) - trypsin , chemistry , polymer , acrylamide , glycidyl methacrylate , succinimide , methacrylate , polymer chemistry , chymotrypsin , copolymer , covalent bond , organic chemistry , enzyme
Reactive polymers have been prepared by copolymeriz‐ing N ‐isopropyl acrylamide (NIPAM) with N ‐acryloxy‐succinimide (NASI) or glycidyl methacrylate (GMA). The amino groups of ligands could react with the residues of NASI or GMA and the polymers could be precipitated by temperature and/or salinity variation, since they contained the NIPAM residues. As a model, p ‐aminobenza‐midine, a trypsin inhibitor, was attached to the polymers to form water‐soluble macroligands, capable of selectively binding trypsin from a trypsin–chymotrypsin solution. After precipitation of the macroligand–trypsin complex, followed by dissociation, approximately 82% trypsin was isolated. The NIPAM‐GMA copolymer was also reacted with immunogammaglobulin (IgG) and alkaline phosphatase (AP). It was demonstrated that the IgG bearing polymer was able to bind protein A and the whole complex was precipitable. The reactive polymer was also used for direct immobilization of AP which was active in repeated reactions.