摘要
用纯化的鸡传染性法氏囊病病毒 ( IBDV) CA毒株和 B87株混合抗原脾内免疫 BALB/C小鼠 ,取其脾细胞与 SP2 /0骨髓瘤细胞融合 ,采用间接 ELISA方法进行筛选 ,共检出 31个原始阳性孔 ,阳性率为 1 5.4%。选择其中 3个孔 ,分别进行 3次克隆 ,最后获得 3株杂交瘤细胞 ,命名为 B2 8、B3 4、C2 8。通过与其它禽源病毒的交叉试验证明 ,分泌的抗体均具有高度特异性。3株细胞的平均染色体数分别为 94、87、89条。3种单克隆抗体均无免疫沉淀特性。相加 ELISA试验结果表明 ,3种单克隆抗体识别的是病毒抗原上互不相同的表位。3株杂交瘤细胞的细胞上清和小鼠腹水单克隆抗体的ELISA效价分别达 1 :1 2 8～ 2 560和 1 0 -3～ 1 0 -6,其中 B3 4显著高于另 2株。通过中和试验证明 ,仅 B3 4单克隆抗体能特异中和 IBDV CA株。连续培养 2 1代 ,B3
Cells of murine myeloma cell line SP/0 were fused with spleen cells of BALB/C mice intrasplenically immunized 6 days before with mixed virus antigen of IBDV strain CA and strain B87.Samples of 31 wells were proved positive by indirect ELISA.Three hybridoma cells,B 24 、B 34 、C 28 , were screened through cloned 3 time separately.These 3 MAbs showing no immunity precipitation reaction were highly specific for IBDV and not reacted with other avian viruses.The ELISA titres of the cell supernates and the ascites were 1∶128~1∶2560 and 10 -3 ～10 -6 respectively,among them,that of the B 34 was the highest.The numbers of chromosomes of these 3 hybridoma cells were 94,87 and 89 respectively.The results of additivily ELISA suggested that these 3 MAbs recognized different epitopes of IBDV.Only Mab B 34 neutralized IBDV strain CA and it secreated MAbs steadily after subcultured 21 times.
出处
《中国兽药杂志》
北大核心
2000年第3期9-13,共5页
Chinese Journal of Veterinary Drug
