Activation of Sepharose with Epichiorohydrin and Subsequent Immobilization of Ligand for Affinity Adsorbent

• Published in: Journal of biochemistry (Tokyo) Vol. 85; no. 4; pp. 1091 - 1098
• Main Authors: MATSUMOTO, Isamu, MIZUNO, Yuko, SENO, Nobuko
• Format: Journal Article
• Language: English
• Published: Oxford University Press 01.04.1979
• ISSN: 0021-924X; 1756-2651
• DOI: 10.1093/oxfordjournals.jbchem.a132417

The optimal conditions for the activation of Sepharose by epichiorohydrin and subsequent immobilization of ligands were investigated. Under the optimal conditions for activation, namely, 30% Sepharose-5 % epichlorohydrin-0·4 M NaOH, 40°C, 2 h, the maximum amount of epoxy group was introduced into Sepharose with low cross-linking. The adsorbents obtained by using N-acetyl-D-glucosamine, tri-N-acetylchitotriose, and glycoprotein as a ligand exhibited no nonspecific adsorption and good permeability for the high molecular substance to be purified, and were stable in an alkaline solution. Solanum tuberosum agglutinin was specifically adsorbed on a tri-N-acetylchitotriose-Sepharose column and was quantitatively recovered by elution with 0·2 M ammonia solution. Furthermore, the column could be repeatedly used under these conditions without reduction of its capacity.