产蛋下降综合征病毒单克隆抗体的制备

Preparation of Monoclonal Antibodies Against EDS-76 Virus

以粗提的ＥＤＳ－７６病毒（ＮＥ－４株）作为抗原，按常规方法免疫ＢＡＬＢ／ｃ鼠，最后一次免疫后第３天取脾细胞与ＳＰ２／０细胞进行融合，采用间接ＥＬＩＳＡ和ＨＩ进行双重筛选，共检出２５个阳性孔，阳性率为１４．８８％。对其中的１１个阳性值较高的孔进行３次克隆，最后获得的１１株杂交瘤细胞，经长期体外培养和冻存后复苏仍能稳定地分泌抗体；ＥＬＩＳＡ阻断试验和与其他病毒的交叉试验证明，其分泌的抗体具有高度特异性。经鉴定，１１株ＭｃＡｂ均为ＩｇＧ，均无免疫沉淀特性，腹水和细胞培养上清的ＥＬＩＳＡ效价分别为１：３２００～１：５１２００和１：１２８～１：２０４８。杂交瘤细胞的染色体数目为８０～１１２，平均９２。尤为重要的是，１１株杂交瘤细胞中有４株分泌抗ＥＤＳ－７６病毒血凝素决定簇的中和单抗，细胞培养上清和腹水的ＨＩ价分别为２１０～２￣（１２）和２￣（２２）～２￣（２４），腹水的中和效价为１：１２８０～１：５１２０。

The myeloma cell line SP2/0 fused with the spleen cells of BALB/c miceconventionally immunized three days early with crude NE-4 virus. Tweny-five positivereactions(up to 14.88 per cent)were screened by indirect ELISA and hemagglutination-inhibition. Of the twenty-five positive wells,eleven hybridoma cell lines steadily secret-ing monoclonal antibody(McAb)highly specific for NE-4 virus by ELISA block andcross tests were screened through three clonings. These eleven monoclonal antibodieswithout precipitation activity were identified as IgG. The ELISA titres of cell super-natant and ascites were 1:128 to 1:2 048 and 1:3 200 to 1:51 200 respectively.Thenumber of chromosome of hybridoma cell lines was 80 to 112,at an average of 92.Es-pecially,there were four out of eleven lines which secreted neutralization antibody a-gainst hemagglutinin determinant of EDS-76 virus. The titres of hemagglutination-inhi-bition of cell supernatant and ascites were2￣(10) to !(212) and 2￣(22)to2￣(24) respectively.