摘要
目的 :研究提取和纯化与人卵透明带 ( HZP)作用更为密切的人精子膜蛋白 ,为改进精子包被抗原的制备方法和筛选避孕疫苗抗原提供科学依据。方法 :1悬浮离心法制备人精子膜蛋白混合溶液 ;2卵子热溶透析法制备 HZP溶液 ;3将人精子膜蛋白混合溶液进行 SDS- PAGE电泳 ,然后全胶电洗脱分别收集蛋白溶液 30管 ;4梅花孔葡聚糖凝胶试验及精卵结合体外实验鉴定出阳性管 ;5以阳性管内的 HZP特异精子膜蛋白溶液包被抗原进行抗精子抗体 ( As Ab)检测。结果 :SDS- PAGE电泳、梅花孔葡聚糖凝胶试验及精卵结合体外实验证实 ,与 HZP结合的人精子膜蛋白有多种成分 ;全胶电洗脱后以第 1 4管内成分之特异性最强 ,与标准蛋白分子量对照 ,其分子量范围为 48～ 5 3k D。结论 :以电洗脱转移法纯化、筛选的人精子膜蛋白是特异的 ,与 HZP具极强相关性 ;以该精子膜蛋白包被抗原用于检测 As Ab,其灵敏度更高 。
Objective:To purify acrosomal membrane protein of sperm for improvement of diagnosis and treatment of immunologic infertility,as well as for selection of antigens for AsAb test and antifertility vaccines.Methods:①Triton digestion was performed on aliquots of human sperm membrane protein collected by centrifugation.②The solubilized human zona pellucida(HZP) was dialyzed against PBS for 48 h with two changes before use.③Thirty tubes were individually collected by SDS PAGE electrophoresis and whole gel eluter system,based on solubilized sperm membrane protein.④The positive tubes containing human sperm membrane proteins that binded HZP were confirmed by gelating agglutination test and sperm zona binding test.⑤AsAb test was performed with coated antigens from the positive tubes.Results:In these procedures,human sperm membrane protein that reacted strongest with the HZP proteins belonged to the molecular region of 48 53 kD in the fourteenth tube,besides weakly reacting proteins in other three tubes among above mentioned 30 tubes.Conclusion:Coating antigens for AsAb test with human sperm membrane proteins in the positive tube will obviously improve diagnosis and treatment of unexplained infertility.
出处
《滨州医学院学报》
2003年第5期326-328,共3页
Journal of Binzhou Medical University
基金
山东省卫生厅资助课题 1996 (AICFB)
关键词
顶体膜蛋白
包被抗原
抗精子抗体
人精子受精相关抗原
人卵透明带
蛋白电泳
避孕疫苗
acrosomal membrane protein
coated antigen
anti sperm antibody
human fertilization related sperm antigens
human zona pellucida
protein electrophoresis
