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Studies on oriented and reversible immobilization of glycoprotein using novel boronate affinity gel
Author(s) -
Liu XiaoChuan,
Scouten William H.
Publication year - 1996
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/(sici)1099-1352(199634/12)9:5/6<462::aid-jmr283>3.0.co;2-g
Subject(s) - chemistry , horseradish peroxidase , agarose , catechol , ligand (biochemistry) , chromatography , affinity chromatography , immobilized enzyme , moiety , boronic acid , combinatorial chemistry , sepharose , covalent bond , enzyme , nuclear chemistry , organic chemistry , biochemistry , receptor
The authors have previously synthesized a novel boronate affinity ligand, catechol [2‐(diethylamino)carbonyl‐4‐bromomethyl]phenylboronate. When this ligand was coupled to cellulose beads, it bound horseradish peroxidase (HRP), a glycoprotein, at pH 7.0. In comparison, commercial m ‐aminophenylboronic acid‐agarose did not bind HRP below pH 8.0. HRP was immobilized in an oriented and reversible fashion using this gel. The immobilized enzyme retained 90.12 per cent of its original activity, probably due to its attachment via the carbohydrate moiety of the enzyme. After repeated use, the activity remaining on the new gel was twice as high as that on conventional m ‐aminophenylboronic acid‐agarose. The column was regenerated easily by washing with dilute acid becuase of reversibility of the boronate glycol bond.