利用已构建的含有HBeAg基因的重组家蚕核多角体病毒rBmHBe ,摸索了HBeAg在家蚕细胞中表达的最佳条件组合 ,细胞接种密度 :(3 .2～ 6 .4)× 10 5细胞 /瓶。病毒接种时间 :细胞接种后 48～ 6 0h。感染复数 (MOI)为 0 .0 4～ 0 .4PFU/细胞。细胞上清经硫铵沉淀、分子筛和阴离子交换柱层析纯化获得电泳纯样品。该样品与E .coli表达的HBeAg相比 ,抗原性高出 10 0倍 ,且与抗HBc交叉反应低至 1%倍。利用该样品建立双抗原夹心法测定抗HBe ,灵敏度高于目前的市售抗HBe试剂盒。 Using the constructed recombinant silkworm polyhedrosis virus rBmHBe containing HBeAg gene, the optimal combination of HBeAg expression in the silkworm cells was explored. The cell seeding density was (3.2-6.4) × 10 5 cells / vial. Virus inoculation time: 48 ~ 60h after cell inoculation. The multiplicity of infection (MOI) was 0. 04 ~ 0. 4 PFU / cell. Cell supernatant was ammonium sulfate precipitation, molecular sieve and anion exchange chromatography purified pure electrophoresis samples were obtained. This sample is 10x more antigenic than E. Coli expressing HBeAg and cross-reacts as low as 1% fold with anti-HBc. The sample was used to establish a double antigen sandwich method for the determination of anti-HBe with higher sensitivity than the current commercial anti-HBe kit.