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抗透明带抗体对透明带溶解、精子穿透以及透明带脱落的抑制效应已被证实,本实验应用细胞融合技术制备猪卵透明带(PZ)单克隆抗体,探讨应用抗PZ 单克隆抗体对PZ 抗原结构及特性进行研究的途径。先后进行了六次细胞融合,五次融合获得成功。共接种1728孔,有克隆生长255孔,平均融合率15%。ELISA 检测246孔,初筛抗体阳性9孔,连续多次检测抗体持续阳性2孔。随后采用有限稀释,进行克隆化培养。卵细胞表面沉淀试验表明,阳性杂交瘤培养上清液与无卵丘卵子表面形成明显的沉淀层。分别采用分子量为1000和4000的PEG 作细胞融合,克隆生长孔各为34与29,阳性克隆孔数分别为2和3。对融合后无饲养细胞培养的观察说明,无饲养细胞生长克隆孔数是13,含饲养层生长克隆孔数为34,融合率各为4.5%与11.8%。
Inhibition of zona pellucida on zona pellucida dissolution, sperm penetration and zona pellucida shedding has been demonstrated. In this study, monoclonal antibodies against porcine zona pellucida (PZ) were prepared by cell fusion technique. The effects of anti-PZ monoclonal antibodies on PZ Antigen structure and characteristics of the way to study. Has carried out six cell fusion, five fusion success. A total of 1728 holes were inoculated, with 255 colonies growing, with an average fusion rate of 15%. 246 wells were detected by ELISA, 9 wells were positive for primary screening, and 2 wells were continuously positive for several consecutive tests. Subsequent use of limited dilution, clonal culture. Egg cell surface sedimentation test showed that positive hybridoma culture supernatant and no cumulus egg surface formed a clear sedimentation layer. PEGs with molecular weights of 1000 and 4000, respectively, were used for cell fusion. The number of clonal growth wells was 34 and 29, respectively. The number of positive clones was 2 and 3, respectively. The observation of the feeder-free culture showed that the number of feeder-free colonies was 13, the number of colonies with feeder layers was 34, and the fusion rates were 4.5% and 11.8% respectively.