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目的制备黄热病毒单克隆抗体并对其生物特性进行鉴定。方法通过动物免疫、细胞融合、阳性克隆筛选等方法制备黄热病毒单克隆抗体,免疫Balb/c小鼠收集腹水,得到扩大制备的单抗,辛酸-饱和硫酸铵法纯化腹水。IFA法检测单抗与黄热病毒(YFV)、乙脑病毒(JEV)、登革病毒1-4型(DENV1-4)、西尼罗病毒(WNV)和基孔肯雅病毒(CHIKV)的特异性反应,并对纯化前后的腹水进行效价分析;ELISA方法进行单抗的亚型分类。结果获得8株黄热病毒的单克隆抗体,均与YFV有特异性反应,1株与JEV有交叉反应,余7株与其它4种虫媒病毒无交叉反应。抗体分型大部分为IgG;纯化后单抗蛋白含量大多在30mg/ml以上。纯化后的4株单抗效价在1∶25600,3株在1∶12800,仅1株效价较低。结论本研究目前获得了几株特异性好、效价高的黄热病毒单克隆抗体,为建立黄热病毒快速检测系统奠定了基础。
Objective To prepare monoclonal antibodies against yellow fever virus and identify its biological characteristics. Methods The monoclonal antibodies of Yellow Fever virus were prepared by animal immunization, cell fusion and positive clonal screening. The ascites were collected from Balb / c mice and the expanded McAbs were obtained. The ascites purified by caprylic acid - saturated ammonium sulfate method was obtained. IFA assay was used to detect monoclonal antibody against YFV, JEV, DENV1-4, WNV and CHIKV Specific reaction, and the titer analysis of ascites before and after purification; ELISA method subtype of monoclonal antibody. Results Eight monoclonal antibodies against yellow fever virus were obtained, all of which reacted specifically with YFV. One strain cross-reacted with JEV and the other seven strains did not cross-react with the other four arboviruses. The majority of antibody typing was IgG; most of the purified monoclonal antibodies were above 30mg / ml. After purification, the titer of the four monoclonal antibodies was 1: 25600, and the number of strains was 1: 12800. Only one titer was lower. Conclusions Several monoclonal antibodies against yellow fever virus with good specificity and high titer were obtained in this study, which laid the foundation for the establishment of rapid detection system of yellow fever virus.