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免疫印迹显示,抗牛精子sp18族膜蛋白的单克隆抗体（sp18单抗）与小鼠精子14,18,22,30和60 ku 5条带有较弱的抗原信号,同时,sp18单抗也能结合鸡卵溶菌酶。间接免疫荧光证实小鼠精子头后部存在sp18抗原。获能2h的小鼠精子与0.182 mg/mL的sp18单抗共孵育15～20min后进行体外受精,结果发现,单抗组的受精率（77.1％）与血清对照组（79.2％）和空白对照组（80.3％）间无显著差异（p>0.05）。继续培养受精卵发现,3组胚胎发育至2-细胞期的比率分别为100％,100％和97.9％（p>0.05）;发育至4-细胞期的比率分别为0,64.1％和64.3％,即单抗组胚胎的发育被完全阻断在2-细胞期（p<0.001）。0.5％链霉蛋白酶消化除去阻断胚胎的透明带后,间接荧光定位发现,2个胚胎细胞表面均存在sp18抗原。0.182mg/mL的sp18单抗与体内受精的胚胎共培养时,胚胎发育至2-细胞期（95.2％vs 92.9％）和4-细胞期（70.5 ％ vs 77.9％）的比例与对照组无显著差异（p>0.05）。结果表明,小鼠精子sp18族膜蛋白在受精时掺入卵母细胞质膜系统,成为胚胎细胞表面具有功能的质膜成分,与突破胚胎发育的2-细胞期阻断现象有关。
Immunoblotting showed that anti-bovine sp18 monoclonal antibody (sp18 monoclonal antibody) had weak antigenic signal with mouse sperm 14, 18, 22, 30 and 60 ku 5, while sp18 monoclonal antibody Can be combined with chicken egg lysozyme. Indirect immunofluorescence confirmed the presence of sp18 antigen in the mouse sperm head. In vitro fertilization was performed after incubating mouse sperm for 2h with sp18 monoclonal antibody (0.182 mg / mL) for 15-20 min. The results showed that the fertilization rate (77.1%) in the monoclonal antibody group was significantly higher than that in the serum control group (79.2% There was no significant difference between groups (80.3%) (p> 0.05). The further development of the fertilized eggs revealed that the rates of embryo development up to the 2-cell stage were 100%, 100% and 97.9%, respectively (p> 0.05), and the rates of development to 4-cell stage were 0,64.1% and 64.3% , That is, the development of the mAb embryos was completely blocked in the 2-cell stage (p <0.001). After 0.5% pronase digestion to remove the zona pellucida that blocks the embryo, indirect fluorescence localization revealed the presence of the sp18 antigen on both embryonic cell surfaces. When the 0.182 mg / mL sp18 McAb was co-cultured with in vivo fertilized embryos, the ratio of embryo development to the 2-cell stage (95.2% vs 92.9%) and 4-cell stage (70.5% vs 77.9%) was not significantly different from that of the control group Difference (p> 0.05). The results showed that sp18 family membrane proteins of mouse sperm were incorporated into the plasma membrane system of the oocyte during fertilization and became the plasma membrane components of the embryonic cell surface, which was related to the phenomenon of 2-cell block during embryonic development.