ABSTRACT Background The hornworn Erinnyis ello is the major pest of natural rubber crops in Colombia, mainly controlled using toxic chemical insecticides. The use of E. ello Betabaculovirus is an environmentally sustainable alternative for its control. The aim of the present work was to characterize a prototype biopesticide formulation and evaluate its efficacy under different conditions. Results Quality control evaluations of formulated biopesticide revealed that all the parameters evaluated were under the permissible level. The lethal concentrations LC 50 and LC 90 of the biopesticide were 4.3 × 10 3 and 5.5 × 10 4 occlusion bodies (OBs) mL −1 , respectively. Biopesticide efficacies against second and fourth instar larvae under greenhouse conditions were higher than 80%. Evaluation of two application rates in a clonal garden resulted in 84% and 88% efficacy, comparable to that obtained with the chemical. The biopesticide in a commercial plantation showed efficacies between 74% and 82%. Biopesticide post‐application persistence was estimated at least in 1 week under field natural conditions. Results allowed selection of the lowest evaluated dose (1 × 10 11 OBs ha −1 ) as the basis for further field evaluations. Conclusion Formulated ErelGV showed high efficacy to control the hornworm in rubber crops and high potential to be included in integrated pest management programs, thus it could be an interesting alternative to replace agrochemicals. © 2018 Society of Chemical Industry