The spore/crystal complex was pre pared from Bacillus thuringiensis var. kurstaki HD-1 and HD-73 during sporulation. The parasporal crystal was isolated from the spore crystal/complex by liquid two phased methods. CryIA and CryIA (c) were precipitated from parasporal crystal on PI4.4. The 130～140 kDa molecular weight protein was separated with sodium-dodecy-sulfate polyacrylamide gel (SDS - PAGE ) electrophoresis. Two polyclonal antibodies were acquired using CrylA , CryIA (c)as antigens. ELISA titer of the two antisera were＞ 1/100000, ＞ 1/100000, respectively. Four monoclonal antibodies ( A4FSF11, B4E6C7, B4E6D8 and B4FSG11) were produced using CryIA insecticidal crystal protein (ICP) to immune BALB/C mouse. A double sandwich ELISA coated with anti-CryIA (c) polyclonal antibody was established with B4E6D8 as sandwich antibody. The toxin level of NUCOTN33B and CCRI-30 in leaf was detected. The content was 23. 4 and 24. 7 ng·g-1 FW,respectively. The result was identical with the insecticidal effect.%用苏云金杆菌菌株HD-1和HD-73分别发酵培养制备了孢晶混合物,孢晶混合物经分离、纯化、电泳得到CryIA、CryIA(c)杀虫晶体蛋白.用CryIA和CryIA(c)分别免疫家兔制备了两种多抗.用CryIA免疫小鼠,经细胞杂交、融合、克隆后筛选出4个单克隆株系A4F5F11、B4E6C7、B4E6D8、B4F5G11.用CryIA(c)多抗包被,B4E6D8上清液作为夹心抗体成功地建立了双抗夹心 ELISA,并检测了中棉所30、NUCOTN33B蕾期叶片中毒蛋白的含量,结果分别为23.4ng·g-1 FW、24.7ng·g-1 FW.
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