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目的:制备嗜水气单胞菌(Aerom onas hydophila)抗独特型单克隆抗体(AIdmAb)并进行鉴定。方法:以具有中和作用的抗嗜水气单胞菌mAb1G10免疫BALB/c小鼠,采用常规杂交瘤技术制备嗜水气单胞菌AIdmAb。用ELISA法对mAb的效价及其模拟嗜水气单胞菌的特性进行鉴定。结果:获得4株能稳定分泌嗜水气单胞菌AIdmAb的杂交瘤细胞株,分别命名为1F9、1H10、3F5、4G3。经测定杂交瘤细胞腹水mAb效价为10-4～10-5。竞争抑制实验结果表明4株AIdmAb均能不同程度地抑制嗜水气单胞菌与兔抗嗜水气单胞菌多克隆抗体结合,其中两株1F9、1H10具有较强的抑制作用。用作制备抗独特性抗体腹水的小鼠,其血清均能检测与嗜水气单胞菌进行免疫结合的抗体,经ELISA检测,其效价为1:100～1:1000。结论:成功地制备了产生嗜水气单胞菌抗独特型抗体的杂交瘤细胞株,为制备该菌的抗独特性抗体疫苗奠定了基础。
Objective: To prepare and identify Aerom onas hydophila anti-idiotypic monoclonal antibody (AIdmAb). Methods: BALB / c mice were immunized with mAb1G10 against A. hydrophila with neutralization effect. Aeromonas hydrophila AIdmAb was prepared by conventional hybridoma technique. The titer of mAb and its characteristics of Aeromonas hydrophila were identified by ELISA. Results: Four hybridoma cell lines stably secreting AIdmAb from A. hydrophila were obtained and named as 1F9, 1H10, 3F5 and 4G3, respectively. The ascites titer of hybridoma cells was 10-4 ~ 10-5. The results of competition inhibition experiment showed that all four AIdmAbs could inhibit the combination of Aeromonas hydrophila and polyclonal antibody against Aeromonas hydrophila to varying degrees, of which two strains of 1F9 and 1H10 had a strong inhibitory effect. The mouse serum used for the preparation of the anti-idiotypic antibody ascites can detect the antibody that binds to Aeromonas hydrophila immunologically, and its titer is 1: 100-1: 1000 by ELISA. Conclusion: The hybridoma cell line producing Aeromonas hydrophila anti-idiotypic antibody has been successfully prepared, which lays the foundation for the preparation of the anti-idiotypic antibody vaccine of this bacterium.