分泌抗登革病毒单抗独特型抗体杂交瘤细胞系的建立  

Establishment of the hybridoma Cell Lines Secreating Antiidiotype Antibodies against the Monoclonal Antibodies of Dengue Virus

作　　者：徐品芳 秦鄂德[1] 司炳银[1] 于曼[1] 杨佩英[1] 阎国珍[1] 

机构地区：[1]军事医学科学院微生物流行病研究所

出　　处：《单克隆抗体通讯》1994年第1期31-35,共5页

基　　金：军事医学科学院基金

摘　　要：将纯化的登革３型病毒（ＤＥＮ－３）单克隆抗体３Ｄ３（ＡｂＩ）连结到载体分子ＫＬＨ上，免疫ＢＡＬＢ／ｃ小鼠，取其脾细胞与Ｓｐ２／０小鼠骨髓瘤细胞融合。用ＥＬＩＳＡ夹心法检测阳性克隆，融合率达１００％，阳性率为５％，经２～３次克隆化，获得了６株分泌抗登革病毒单抗的独特型抗体（ＭｃＡｂ２）的杂交瘤细胞系。在进行阳性筛选及特异性鉴定时，选择了以３～５μｇ／ｍｌ的最适Ａｂ１包被浓度，并用正常小鼠Ｉｇ及ＫＬＨ分别包被反应板作对照，以排除其抗异种免疫球蛋白的干扰。The purified monoclonal antibody(Ab1)3D3 against dengue type 3 virus was coupled chemically to carrier protein-Keyhole limpet hemocyanin(KLH).The conjugates of Ab1-KLH were injected into BALB/c mice.The spleen cells from this animal were fused with mouse myeloma Sp2/0 cells. The positive clones of antibodysecreting hybridomas were tested by using a sandwich ELISA method.The hybrid efficiency for the fusion was approximately 100%, and the percentage of the hybridomas demonstrating anti-idiotype activity was 5%.Through three times cloning,six hybridoma cells secreting anti-idiotypic antibodies(McAb2)against Ab1 were obtained.After repeated frozen storage in liquid nitrogen and revival,the three hybridoma cell lines can steadily produce high titers of McAb2.When the positive clones and specificity of hybridoma oells were identified by using the ELISA method,0.3-0.5μg Ab1/well coated was optimum.At the same time, the wells of plate were coated with normal mouse Ig and KLH as negative control to rule out a possible interference of the heteroantibody.The idiotypes in the monoclonal antibody molecules of the other type dengue viruses were measured with the McAb2 against Ab1 3D3.

关 键 词：杂交瘤 单克隆抗体 登革热病毒 

分 类 号：R373.33[医药卫生—病原生物学]