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用纯化的36kD蛋白免疫Balb/c小鼠。取免疫脾细胞与小鼠骨髓瘤细胞SP2/0以PEG进行细胞融合,融合率为86％。采用ELISA筛选出杂交瘤细胞阳性孔。经过4～7次有限稀释克隆化,共获得7株能稳定分泌36kD相应MCAb的杂交瘤细胞。将杂交瘤细胞分别注射小鼠腹腔,测定腹水McAb的效价为1:5×10~3～1:1×10~5,抗体亚型均为IgG1。用荧光显微镜观察,结果发现EC2作用的人精子头颈部出现明显荧光反应,提示共同抗原存在于人精子头颈部。这些结果有助于进一步探讨细菌感染引起男性不育的机制以及研制细菌避孕疫苗等工作。
Balb / c mice were immunized with purified 36 kD protein. Immunosuppressive spleen cells and mouse myeloma cells SP2 / 0 with PEG for cell fusion, the fusion rate was 86%. The positive wells of hybridoma cells were screened by ELISA. After 4 to 7 times limiting dilution cloning, obtained a total of 7 strains can stably secrete 36kD corresponding MCAb hybridoma cells. The hybridoma cells were respectively injected into the peritoneal cavity of mouse and the titer of ascites McAb was 1: 5 × 10 ~ 3 ~ 1: 1 × 10 ~ 5. The antibody subtypes were both IgG1. Fluorescence microscopy showed that there was a significant fluorescence reaction in the head and neck of human spermatozoa EC2, suggesting that common antigens exist in the human sperm head and neck. These results help to further explore the mechanism of bacterial infection caused by male infertility and the development of bacterial contraceptive vaccine work.