水蛭素的亲和层析
水蛭素的亲和层析(任务书,开题报告,外文翻译,论文9000字)
摘要
水蛭是一种罕见的中草药。被称为水蛭素的水蛭分泌物是世界上最强的凝血酶抑制剂。天然水蛭素主要分泌于医用水蛭的唾液腺，多半的水蛭素制备方法基于全蛭。水蛭体内含有较多的与水蛭素结构相似的伪成分。这些成分的存在会大大增加分离和纯化水蛭素的难度。根据水蛭素在水蛭身体各个部位的含量的不同，分别对水蛭的嘴部，头部和其他身体部位进行提取。这么做将大大提高生产效率，从而为水蛭素的药用提供了可能。 同时分别采用猪凝血酶，牛凝血酶，人凝血酶三种酶制备亲和层析柱，选择出分离效率最高的层析柱。实验表明，人凝血酶亲和层析柱柱效最好。
关键词：水蛭 凝血酶 水蛭素 分离 亲和层析
Separation of hirudin by affinity chromatography
Abstract
Minamata is a rare Chinese herbal medicine,and its secreted sputum contains an anticoagulant substance,which is known as hirudin. Hirudin is the world’ s most powerful inhibitor of thrombin. Natural hirudin is mainly produced by saliva glands of medical leeches. Most of the methods for preparing hirudin are based on whole leeches. The body of blisters has a large amount of pseudo hirudin. Its structure is similar to that of hirudin,but it has no biological activity. The existence of pseudo- hirudin will increase the difficulty of separating and purifying hirudin. This experiment will divide the body of the blister into three parts and extract the mouth,head and other parts of the blister,so that the separation pseudo The unnecessary troubles caused by hirudin,which greatly increase production efficiency,provide possibilities for the medicinal use of hirudin. At the same time,the affinity chromatography column was prepared using three enzymes:porcine thrombin,bovine thrombin and human thrombin,and the column with the highest separation efficiency was selected.Experiments show that the human thrombin affinity column has the best efficiency. [资料来源：http://www.doc163.com]
Keywords:Leech;Thrombin;Hirudin;Separation;Affinity chromatography
[资料来源：www.doc163.com]
目录
摘要…………………………………………………………………………………I
ABSTRACT…………………………………………………………………………II
第一章 绪论……………………………………………………………………… 1 [资料来源：http://Doc163.com]
1.1水蛭素提纯现有方法介绍…………………………………………………1
1.2水蛭素的结构与特性………………………………………………………2
1.3研究水蛭素亲和层析的目的和意义………………………………………3
1.4水蛭素的应用前景…………………………………………………………3
第二章 试验方法…………………………………………………………………6
2.1 实验材料、试剂、仪器……………………………………………………6 [资料来源：http://Doc163.com]
2.2水蛭的预处理………………………………………………………………7
2.3水蛭素的初步粗提取………………………………………………………7
2.4阴离子交换层析柱的制备………………………………………………7
2.5制备凝血酶亲和层析柱…………………………………………7
2.6水蛭素的进一步分离提纯………………………………………………9
2.7提取物生物活性的测定…………………………………………9
第三章 结果与分析………………………………………………11
3.1丙酮浓盐酸浸提结果……………………………………………11
3.2洗脱剂最适pH的确定…………………………………………………11
3.3滴定结果分析……………………………………………………12
第四章 总结与展望……………………………………………………………14
结语…………………………………………………………………………15 [来源：http://Doc163.com]
参考文献…………………………………………………………………………16
致谢…………………………………………………………………………………18
[资料来源：http://doc163.com]