抗兔附睾特异性蛋白ESP－2单克隆抗体及其抗生育研究

兔附睾液中分离纯化了两个特异性蛋白ＥＳＰ－１和２，分子量分别为４２ＫＤ和２０ＫＤ。应用基因克隆技术，又从人睾丸λ－ｇｔｌｌ表达性。ＤＮＡ基因库中筛选分离出一个１．９Ｋｂ的编码ＥＳＰ－２ｃＤＮＡ片段，为了进一步深入研究ＥＳＰ－２做为抗生育疫苗的可能性，本研究用半固体融合细胞培养基法成功地制备了抗ＥＳＰ－２的单克隆抗体，经三轮筛选后获得阳性克隆３８个，经定量分析ＯＤ值均为０．５以上，其中三株命名为ＥＳＰ－２ｍＡｌ、ＥＳＰ－２ｍＡ３和ＥＳＰ－２ｍＡ９，接种于ＢＡＬＢ／Ｃ小鼠腹腔之后，腹水滴度均达１∶１００，０００。本研究采用体内外结合的被动免疫抗生育实验，即用抗体处理精子后进行体内抗生育实验，用小鼠双侧子宫互为对照和实验，证明ＥＳＰ－２ｍＡ３，ＥＳＰ－２ｍＡ９单抗对小鼠受精有明显的抑制作用，Ｐ＜０．０５和＜０．０１。对上述两株ＥＳＰ－２抗体的抗受精机理需进一步探讨。

The study of Antifertility Effect of Monoclonal Antibodies toThe Epididymis Protein of Rabbit

The rabbit epididymal fluid proteins(ESP-1,ESP-2) have been isolated and purified,42KDa and 20KDa for molecular weights,respectively.An ESP-2 cDNA fragment(1.9kb)encoding ESP-2 protein was screened from human testis expression cDNA libraryλ-gtll with geneclonal technique.In the present study,monoclonal antibodies to the specific rabbit epididymis protein (ESP-2) have been prepared with semi-solid cell culture medium in order to further studywhether it is possible for ESP-2to be an antifertility vaccine.38 monoclonal antibodies have beenobtained through3turn-slelections.Quantitative analysis demonstrated that the optical density(OD)values of monoclonal antibodies concentrations in culture media were more than 0.5 ODvalue.Three monoclonal antibodies were named ESP-2mAl,ESP-2mA3,ESP-2mA9,respectively.Then they were inoculated into BALB/C mice with intraperitoneal injection.The titers ofantibodies in ascites were more than 1:100,000.An in vivo inactive immuno anti fertility test wascarried out.The results showed that monoclonal antibodies ESP-2mA3,ESP-2mA9 inhibitedinsemination of mice (P<0.05,P<0.01).The mechanism of insemination inhibition for ESP 2mA3 and ESP-2mA9 needs to be investigated.