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用热溶猪卵透明带抗原免疫BALB/C 小鼠,取其脾细胞和SP2/O 细胞融合产生的杂交瘤。用间接免疫荧光和ELISA 方法筛选,经四次克隆化后,获得三株分泌猪卵透明带单克隆抗体杂交瘤细胞株(LPD_8,LPC_4,LPD_2)。通过间接免疫荧光试验,其中二株单克隆抗体(LPD_8,LPC_4)与人卵透明带有交叉反应。但它们在猪与人卵透明带上呈现的荧光部位有明显的不同,LPD_8位于整个透明带上,而LPC_4只见于透明带外层。LPD_2荧光位于透明带内层且与人卵无交叉反应。我们认为这种差别是由于抗原决定簇的不同所致。
BALB / C mice were immunized with zona pellucida antigen and hybridomas from spleen cells and SP2 / O cells were obtained. After screening by indirect immunofluorescence and ELISA, three hybridoma cell lines (LPD_8, LPC_4, LPD_2) secreting porcine zona pellucida were obtained after four clones. Through indirect immunofluorescence assay, two monoclonal antibodies (LPD_8 and LPC_4) cross-reacted with human zona pellucida. However, there is a clear difference between the fluorescent parts of porcine zona pellucida. LPD_8 is located on the whole zona pellucida and LPC_4 is only found on the outer part of zona pellucida. LPD_2 fluorescence is located in the inner zona pellucida and no cross-reaction with human eggs. We think this difference is due to the different antigenic determinants.