Analysis of sperm antigens by sodium dodecyl sulfate gel/protein blot radioimmunobinding method.
Analysis of sperm antigens by sodium dodecyl sulfate gel/protein blot radioimmunobinding method.
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通过十二烷基硫酸钠凝胶/蛋白质印迹放射免疫结合法分析精子抗原。
DOI：
10.1016/0003-2697(82)90617-0
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发表时间：
1982
影响因子：
2.9
通讯作者：
A. Menge
中科院分区：
文献类型：
--
作者：
C. Lee;Y. S. Huang;P. Hu;V. Gomel;A. Menge
本文建立了一种基于十二烷基硫酸钠凝胶上复性蛋白质在硝酸纤维素滤纸上印迹的放射免疫结合方法，用于分析引起血清抗精子抗体的精子抗原。用同种附睾精子与完全弗氏佐剂混合免疫家兔，产生抗精子抗体。在十二烷基硫酸钠凝胶上，雄性或雌性家兔的抗血清与三条主要的精子蛋白带反应，相应的分子量分别为70，000 ± 5000、14，000和13，000。在人类中，针对人类精子的单克隆抗体通过杂交瘤技术产生。在产生的六个独立的杂交细胞系中，其中三个显示出分泌免疫球蛋白，其与十二烷基硫酸钠凝胶上的相同的两个蛋白条带反应，其具有大约10，000的分子量。同样的程序也用于分析通过已知技术显示含有抗精子抗体的人血清样品。在人类中引发抗精子抗体的独特精子抗原被鉴定并关联。本研究的结果表明，十二烷基硫酸钠凝胶/蛋白质印迹放射免疫结合法可能是一个敏感的和有用的工具，用于研究精子抗原，引起自身免疫反应及其与人类不育症。
A radioimmunobinding method based on the blotting of renatured proteins from sodium dodecyl sulfate gels on to nitrocellulose filter papers was developed to analyze the sperm antigens that elicit serum anti-sperm antibodies. In rabbits, serum anti-sperm antibodies were raised by immunization with homologous epididymal spermatozoa mixed with complete Freund's adjuvant. The raised antisera from either male or female rabbits were shown to react with three major sperm protein bands on sodium dodecyl sulfate gels with the corresponding molecular weights of about 70,000 ± 5000, 14,000, and 13,000, respectively. In humans, the monoclonal antibodies against human sperm were raised by a hybridoma technique. Out of six independent hybrid cell lines that were generated, three of them were shown to secrete immunoglobulins that react with the same two protein bands on sodium dodecyl sulfate gels, which have the approximate molecular weight of 10,000. The same procedure was also used to analyze human serum samples that were shown to contain anti-sperm antibodies by the known techniques. Unique sperm antigens that elicit anti-sperm antibodies in humans were identified and correlated. The results of this study suggest that sodium dodecyl sulfate gel/protein blot radioimmunobinding method may be a sensitive and useful tool for the study of sperm antigens that elicit autoimmune responses and their association with human infertility.