抗脊髓灰质炎病毒Ⅱ型Sabin株D抗原单克隆抗体的研制  

Preparation and identification of monoclonal antibody against D-antigen of poliovirus type Ⅱ Sabin strain

作　　者：褚学者[1] 姜述德[1] 孙明波[1] 周铁群[2] 方捍华[2] 杨建勇[3] 张新文[1] 潘玥[1] 李卫东[1] 廖国阳[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所IPV课题组,昆明650118 [2]中国药品生物制品检定所疫苗三室,北京100050 [3]北京生物制品研究所菌苗室,北京100024

出　　处：《免疫学杂志》2006年第4期387-389,共3页Immunological Journal

基　　金：云南省科技厅新药专项(2000XY05);2003年度云南省科技厅创新人才引培项目(2003PY07)资助

摘　　要：目的应用杂交瘤技术制备抗脊髓灰质炎病毒Ⅱ型Sabin株D抗原的单克隆抗体。方法用脊髓灰质炎病毒Ⅱ型Sabin株D抗原免疫雌性Balbc小鼠,取其致敏脾脏淋巴细胞与SP20小鼠骨髓瘤细胞融合。间接ELISA法筛选阳性克隆,制备腹水。检测McAb效价、抗体亚类和杂交瘤细胞核型。结果获得4株阳性杂瘤细胞株,选择一株制备腹水,ELISA效价为1∶2.4×106,中和试验效价1∶1.3×104,蛋白浓度为27.2mgmL。McAb与Ⅱ型Sabin株C抗原(ⅡC)、Ⅰ型Sabin株及Ⅲ型Phizer株D抗原(ⅠD、ⅢD)不发生交叉反应,具有高度特异性。结论成功的制备了针对脊髓灰质炎病毒Ⅱ型Sabin株D抗原的McAb,为进一步研究IPV效力检测奠定了基础。Objective To prepare and identify monoclonal antibody （McAb） against the D-antigen of poliovirus type H Sabin strain. Methods Monoclonal antibodies against D-antigen of type Ⅱ Sabin strain were prepared by hybridoma technique. The specificity and titer of the prepared McAbs were evaluated by indirect ELISA and microneutralization analysis. Results Four hybridomas secreting McAb against the D-antigen of poliovirus type H Sabin strain were selected out. The ascitic antibody titers measured with ELISA and nerutrahzation assay were 1 ： 2.4× 10^6 and 1：1.3× 10^4, respectively. The McAb could react with the D-antigen of poliovirus type H Sabin strain, but not with C-antigen of poliovirus type H Sabin strain. Condusion Four hybridoma cell lines secreting McAb specific against the D-antigen of poliovirus type H Sabin strain are obtained.

关 键 词：脊髓灰质炎病毒Ⅱ型Sabin株 单克隆抗体 D抗原 

分 类 号：R392[医药卫生—免疫学]