Preparation of three types of heparin-Sepharose and their binding activities to thrombin and antithrombin III

✍ Scribed by Miyuki Funahashi; Isamu Matsumoto; Nobuko Seno

• Book ID: 102985877
• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 634 KB
• Volume: 126
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90537-1

✦ Synopsis

Three types of immobilized heparin, heparin-Sepharoses I, II, and III, were prepared to study the effect of the mode of the heparin attachment on the binding activities to thrombin and antithrombin III (AT-III). Heparin-Sepharose I was prepared by the coupling of the car-boxy1 groups of heparin with amino-Sepharose, obtained by the epoxy activation of Sepharose 4B with epichlorohydrin and subsequent amination, with the aid of water-soluble carbodiimide. Heparin-Sepharose II was prepared similarly by the coupling of the amino groups of heparin with the carboxyl groups of carboxyl-Sepharose obtained by the succinylation of amino-Sepharose. Heparin-Sepharose III was prepared by the reductive amination of heparin and amino-Sepharose. with sodium cyanoborohydride. Heparin-Sepharoses I, II, and III contained heparin in the ratio of 16: 1:3. However, their binding activities to thrombin and AT-III were not proportional to the heparin content, and heparin-Sepharose III was found to be the most efficient affinity adsorbent for both proteins.