Shigeta M, Takada Y, Ikuma K, Kubota K, Koyama K, Isojima S
Nihon Sanka Fujinka Gakkai Zasshi. 1985 Oct;37(10):2031-8.
Rat spleen cells immunized to human azoospermic semen were fused with mouse myeloma cells (P3 X 63Ag8Ul) to obtain hybridomas which produce monoclonal sperm immobilizing antibodies. Of 114 hybrid clones obtained, 10 were found to produce antibodies with sperm immobilizing activity with complement. One clone (1C4) producing the highest antibody titer was chosen for further characterization. Chromosome analysis showed that 1C4 hybridoma cells contained two types of chromosomes which were derived from rat and mouse. Rat Ig was detected in both the cytoplasma and the culture medium of the cells by direct immunofluorescent test and immunodiffusion test, respectively. Gammaglobulin fractions from the culture medium had a strong sperm immobilizing activity and showed a common single precipitin line against human seminal plasma and human milk proteins. It was proved that the monoclonal antibody was produced to No. 7 antigen and not to No. 3 antigen (lactoferrin) in human seminal plasma, both of which have a common antigenicity to human milk proteins and compose the sperm coating antigens.
将经人无精子症精液免疫的大鼠脾细胞与小鼠骨髓瘤细胞（P3 X 63Ag8Ul）融合，以获得产生单克隆精子制动抗体的杂交瘤。在获得的114个杂交克隆中，发现10个产生具有补体介导精子制动活性的抗体。选择产生最高抗体滴度的一个克隆（1C4）进行进一步鉴定。染色体分析表明，1C4杂交瘤细胞含有两种分别来源于大鼠和小鼠的染色体。通过直接免疫荧光试验和免疫扩散试验分别在细胞的细胞质和培养基中检测到大鼠Ig。培养基中的γ球蛋白组分具有很强的精子制动活性，并且针对人精浆和人乳蛋白显示出一条共同的单一沉淀线。已证明该单克隆抗体针对人精浆中的7号抗原产生，而非3号抗原（乳铁蛋白），这两种抗原与人乳蛋白具有共同抗原性，且构成精子包被抗原。
