IBDV单克隆抗体杂交瘤细胞株的建立及其应用  

The Construction of Hybridomas Secreting Monoclonal Antibodies against Infectious Bursal Disease Virus in Chicken and Its Application

作　　者：乔忠[1] 詹丽娥[1] 史振心 宁官宝[2] 

机构地区：[1]山西省农业科学院畜牧兽医研究所 [2]山西农业大学

出　　处：《华北农学报》1990年第1期98-104,共7页Acta Agriculturae Boreali-Sinica

摘　　要：用经硫酸铵沉淀、高速离心并用蔗糖密度梯度离心提纯的鸡传染性法氏囊病病毒(IBDV)免疫BALB/C小鼠.取免疫鼠脾细胞与NS—1小鼠骨髓瘤细胞触合,共获11株阳性杂交瘤细胞.经3次克隆化和ELISA检测,筛选出3个(1B_1、5D_6、6D_8)能持续分泌抗IBDV单克隆抗体的杂交瘤细胞株.细胞上清液的ELISA效价为1B_1,3.2×10^(-2);5D_6,6.4×10^(-2);6D_8,3.2×10^(-2).腹水效价为1B_1,1.6×10^(-5);5D_6,8×10^(-4);6D_8,4×10^(-3).3株杂交瘤细胞的染色体数分别为105(1B_1),97(5D_6),85(6D_8).它们所产生的单抗皆为IgG1,所对抗的抗原决定簇不相同.3株McAb皆有沉淀特性.利用间接ELISA抑制试验,在诊断中进行了应用.BALB/C mice were immunized with infectious bursal disease virus(IBDV) which had been purified with ammonium sulfate precipitation, high-speed centrifugation and ultracentrifugation in a sucrose gradient.From the fusion of myeloma cells NS-1 with immun spleen cells, 11 strains of positive hybridoma cells occurred.After cloning three times and ELISA test, three strains of hybridomas (1B1, 5D6, 6D8) were selected which were able to produce monoclonal antibodies to the IBDV continuously. Cell supernatant valences are 1B1, 3.2×l02; 5D6, 6.4×102; and 6D8, 8×l02; Ascites fluid valances are 1B1, 1.6×l05; 5D6, 8×l04; and 6D8, 3.2×103, Their chromosome are 1B1 105, 5D6 97 and 6D8 85. IBDV McAbs didn't cross-react with ND,MD and IA antigens. Their immunoglobulin class/subclass are all IgG1.They have different antigenic determinants and all show a characteristic of precipitation. The primary application of the indirect ELISA inhibiting assay for detection of IBDV antibodies in chicken is investigated.

关 键 词：鸡 法氏襄病毒 单克隆抗体 

分 类 号：S858.310.5[农业科学—临床兽医学]