摘    要 毒蛇咬伤是一种突发性的急性中毒事件，病情变化快，症状凶险复杂。随着抗蛇毒药物不断发展，抗蛇毒血清成为治疗毒蛇咬伤的特异性药物，提高了蛇伤救治水平。抗蛇毒血清以蛇毒类毒素为抗原，辅以免疫佐剂给实验动物反复进行注射免疫，动物机体受刺激后引起免疫细胞产生应答，最终由浆细胞合成具有与相应蛇毒特异性结合并能中和其毒性的免疫球蛋白IgG。抗蛇毒血清的疗效，无论是动物试验还是临床疗效，是世界公认的最有效的药物。蛇毒成分种内差异，表明利用混合蛇毒作为抗蛇毒血清生产免疫源的重要性。本研究以实验室自制菜花原矛头蝮（Protobothrops jerdonii)、原矛头蝮（Protobothrops mucrosquamatus）和乡城原矛头蝮（Protobothrops xiangchengensis）三种蛇毒混合样品所制备的类毒素对大白兔皮下多点免疫注射制备的多价抗蛇毒血清，与商用抗蝮蛇毒血清为对象，用抗体组学技术,去除抗蛇毒血清中与目的抗体不相关的成分，通过RP-HPLC对抗体效价进行评估。实验结果表明自制的多价抗蛇毒血清对于蝮蛇毒中毒患者没有商用血清的治疗效果好，但采用RP-HPLC评估抗体效价的方法是可行的。90192

Abstract Snake bites is a sudden acute poisoning events, the condition changes quickly, the symptoms are dangerous complex。 With the anti-snake venom drugs continue to develop, antivenom has become a specific drug for treatment of snakebite, improved the level of snake injury treatment。 Anti-snake venom serum to snake venom toxin as antigen, supplemented by immune adjuvant to the experimental animals repeated immunization, animal body stimulation caused by immune cells to produce a response, and ultimately the plasma cell synthesis immunoglobulin IgG which can be specifically combined with the snake venom and neutralize its toxic。 The curative effect of antivenoms, both from animal experiments and clinical curative effect, is the most effective drugs recognized all around the world。 Snake venom component intraspecific differences, indicates the importance of using the mixture of snake venom as antivenin to produce immune source。 This research used antiserum from commercial and laboratory homemade by subcutaneous immunized injecting the toxoid prepared by mixing three kinds of venom samples: Protobothrops jerdonii、Protobothrops mucrosquamatus and Protobothrops xiangchengensis into rabbits to prepare the anti-snake venom serum。 Remove the components not related to the target antibody in anti-snake venom serum by antibody omics technique, assess the snake venom serum titer by RP-HPLC。 Experimental results show that the multivariate antivenom serum homemade is not effective for the treatment of commercial anti-serum in patients with Protobothrops venom poisoning, but it is feasible to use RP-HPLC to evaluate antibody titers。

毕业论文关键词：蛇毒；抗蛇毒血清；效价；反相-高效液相色谱

Keyword: snake venom；antivenom；titer；RP-HPLC

目    录

引言 4

1 材料与方法 5

1。1实验材料 5

1。2实验方法 5

1。2。1 实验室自制抗蛇毒血清 5

1。2。1。1类毒素抗原制备及兔子免疫 5

1。2。1。2抗蛇毒血清的采集 5

1。2。2免疫球的蛋白的源Q于W优E尔A论S文R网wwW.yOueRw.com 原文+QQ75201,8766 分离纯化 6

1。2。3抗体亲和层析柱的制备 6

1。2。4蛇毒的亲和吸附、冻干