家蚕微粒子孢子单克隆抗体的研制及其在检测上的初步应用

将家蚕微粒子Nosema bombycis孢子免疫的BALB/c小鼠的脾细胞和FO细胞融合,经克隆筛选获得6株能稳定分泌抗N.bombycis孢子表面抗原的单克隆抗体细胞株(Nb_(1-6))。其中Nb_(1-2)和Nb4分泌的抗体属于IgM,Nb_3,Nb_5和Nb_6分泌的分别为IgG_2b、IgG和IgG_3。特异性检查结果表明,除Nb_4和Nb_6对柞蚕微粒子孢子有交叉凝集反应外,其余均对家蚕N.bombycisJ孢子具特异性。杂交瘤细胞培养上清液对N.bombycis孢子的凝集效价以Nb_1为最高,达1:256,由此细胞株诱发的腹水效价高达1:5000。建立了以普通聚苯乙烯酶标板为载体,完整的N.bombycis孢子为抗原的LEISA测定方法,最低检测量为700个孢子。

PRODUCTION OF MONOCLONAL ANTIBODIES TO PEBRINE PROTOZOA OF DOMESTIC SILKWORM AND THEIR APPLICATION

Monoclonal antibodies ( Mabs ) to Nosema bombycis of silkworm were generated by PEG fusion of splenocytes from immunized BALB/c mice and FO myeloma cells. After extensive screening, 6 hybridoma cell strains (Nb1-6)were selected which secrete Mabs to antigens on the surface of N.bombycis. IgM was secreted by Nb1-2 and Nb4, IgG2b, IgG1, and IgG3 were secreted by Nb3,Nb5 and Nb6 respectively. With the exception of Nb4 and Nb6 which are cross-reacting antibodies to Nosema sp.of Antheraea pcrnyi and to N. bombycis, every strain is idiosyncrasy to N.bombycis. Nb1 produced the highest titer of supernatant and of ascites. The agglutination reaction of the former was arornd 1:256 and that of the latter around 1:5 000. ELISA method using ordinary polystyrene as carrier and complete spores of N. bombycis as antigen was established, and the sensitivity amounted to 700 spores.