肝素-琼脂糖凝胶6FF的制备及其在抗凝血酶III分离纯化中的应用

Preparation of Heparin-Agarose Gel 6FF and Its Application in the Purification of Antithrombin III

以自制琼脂糖凝胶6FF为基质,肝素为配基,利用还原胺化方法制备了肝素-琼脂糖凝胶6FF介质.考察了肝素偶联反应的影响因素,并对其进行了优化,得到肝素配基的含量为2.7mg/mL.用所制介质从人血浆中分离出抗凝血酶III,从血浆开始计算的抗凝血酶III活性回收率为31.76%,与商品Heparin-SepharoseCL-6B的分离效果相当.

The self-prepared bead-formed agarose gel was used as the affinity support, and heparin was employed as the ligand. Heparin was linked covalently to amino-agarose beads, which was prepared by amination of epoxy-agarose beads. The effects of operating conditions on the coupling reaction were investigated and optimized. The amount of heparin immobilized on the synthesized gel was 2.7 mg/mL medium. The synthesized affinity medium was used to isolate antithrombin Ⅲ from human plasma. The recovery of antithrombin Ⅲ activity was 31.76%, which was similar to that of Heparin-Sepharose CL-6B from Pharmacia, Sweden.