Insect cell-baculovirus expression system has been widely used in insect pesticide and recombinant protein production on a large scale.The interplay among the multiplicity of infection (MOI)，time of infection (TOI) and initial cell density (ICD) in insect cell-recombinant expression systems is not clearly understood and the literature related to the investigation of the quantitative relationship of MOI，TOI and ICD is very scarce.In the present study，an orthogonal design was used to select the optimum conditions，in which the effects of MOI，ICD and TOI on the budded virus (BV) titer and occluded virus (OV) number were evaluated in the HzAM1 insect cell-recombinant baculovirus system at a low MOI in shake-flasks.The results indicated that among MOI，TOI and ICD，MOI had a significant influence on the amounts of BVs and OVs produced.The optimal conditions for the production of both BVs and OVs in the HzAM1 insect cell-recombinant baculovirus system were：MOI of 0.1，initial cell density of 2×105 cells·ml-1 and infection at the early exponential phase.A maximum BV concentration of 2.0×107 TCID50·ml-1，was obtained.Furthermore，the optimal condition obtained was applied to the scale-up test in the airlift reactor，and a maximum BV concentration of 2.13×107 TCID50·ml-1 was obtained，which was bigger than those in the shaked flask.