肝素-壳聚糖亲和层析纯化猪凝血酶研究  

Purification of porcine thrombin by affinity chromatography of heparin-chitosan

作　　者：何晓梅[1,2] 韦传宝[1,2] 张业峰[1] 

机构地区：[1]皖西学院生物与制药工程学院,安徽六安237012 [2]皖西学院安徽省植物生物技术实验实训中心,安徽六安237012

出　　处：《食品工业科技》2013年第4期176-179,183,共5页Science and Technology of Food Industry

基　　金：安徽省教育厅自然科学基金重点项目资助(KJ2010A328;KJ2011A272)

摘　　要：采用反相悬浮交联法制备壳聚糖球珠,壳聚糖球珠经过环氧氯丙烷活化并与肝素耦联制成亲和层析剂分离和纯化猪凝血酶。结果表明制备出几何形态、分散性、溶胀性均良好的壳聚糖球珠的最佳条件是:5%壳聚糖溶液112.5mL、液体石蜡75mL、乙酸乙酯15mL、甲醛10mL、戊二醛2mL、Span808滴、转速2500r/min。壳聚糖球珠用ddH2O溶胀24h,具有明显的多孔性。猪凝血酶活性检测结果表明肝素-壳聚糖亲和层析剂能够用于猪凝血酶的分离和纯化。此研究结果对于亲和层析剂的制备和猪凝血酶的分离和纯化具有一定的参考价值。Chitosan beads were prepared by inverse suspension crosslinked polymerization. Chitosan beads, activated by epichlorohydrin,that were coupled with heparin to prepare affinity chromatography reagent. The prepared affinity chromatography reagent of heparin-chitosan beads was used to purify porcine thrombin. Results showed that optimum parameters of preparing chitosan beads were 112.5mL of 5% chitosan solution, 75mL of liquid paraffin, 15mL of acetic ether, 10mL of formaldehyde,2mL of glutaraldehyde,8 drops of Span 80 and stirring speed of 2500r/min. After swelling with ddH20 for 24h,the prepared chitosan beads showed apparent porosity. Detection results of porcine thrombin activity confirmed that the affinity chromatography of heparin-chitosan beads could be used in the purification of porcine thrombin. These results could present some references for the preparation of affinity chromatography reagent and the purification of porcine thrombin.

关 键 词：壳聚糖球珠 肝素 亲和层析 纯化 凝血酶 

分 类 号：Q556.3[生物学—生物化学]