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目的 :建立抗基孔肯亚病毒单克隆抗体细胞株 ,筛选具有中和活性的特异性单克隆抗体。方法 :利用细胞融合技术建立抗基孔肯亚病毒单克隆抗体杂交瘤细胞株 ,用细胞培养中和试验初筛具有中和活性的单克隆抗体 ,用固定单克隆抗体稀释病毒方法进行乳鼠中和试验 ,进一步验证单克隆抗体的保护作用 ;用中和交叉试验鉴定中和单克隆抗体反应的特异性。结果 :用免疫荧光法检测建立了 9株能稳定分泌抗基孔肯亚病毒单克隆抗体的杂交瘤细胞株 ,初筛出 4株具有中和活性的单克隆抗体 ,其中 1F1单克隆抗体稀释 2 0 0倍时可保护 50 %细胞不产生细胞病变 ;乳鼠中和试验1F1单克隆抗体中和指数为 10 3.3,对试验小鼠有较强的保护作用。中和交叉试验表明 ,1F1单克隆抗体只中和基孔肯亚病毒 ,与其他病毒无交叉反应 ,特异性较高。结论 :制备了 9株抗基孔肯亚病毒单克隆抗体杂交瘤细胞株 ,筛选得到了 1F1具有较强中和活性的特异性单克隆抗体 ,可望为基孔肯亚病毒病的诊断、紧急预防与治疗提供良好试剂
OBJECTIVE: To establish a monoclonal antibody against Chikungunya virus and screen for specific monoclonal antibodies with neutralizing activity. Methods: The hybridoma cell line of monoclonal antibody against Chikungunya virus was established by cell fusion technique. The monoclonal antibody with neutralizing activity was screened by cell culture neutralization test and the virus was diluted with immobilized monoclonal antibody And tests to further verify the protective effect of the monoclonal antibody; neutralization and cross-tests were used to identify the specificity of the neutralizing monoclonal antibody response. RESULTS: Nine hybridoma cell lines stably secreting monoclonal antibodies against Chikungunya virus were established by immunofluorescence assay. Four monoclonal antibodies with neutralizing activity were initially screened, in which 1F1 monoclonal antibody was diluted 2 0 0 times can protect 50% of cells does not produce cytopathic; Neutralization test 1F1 monoclonal antibody neutralization index of 10 3.3, the test mice have a strong protective effect. The neutralization crossover test showed that the monoclonal antibody 1F1 neutralizes only Chikungunya virus and has no specific cross-reactivity with other viruses. CONCLUSIONS: Nine hybridoma cell lines producing monoclonal antibodies against Chikungunya virus were prepared. The specific monoclonal antibodies against 1F1 with strong neutralizing activity were screened, which is expected to be the diagnosis of chikungunya virus disease. Prevention and treatment to provide good reagents