title: Expression and purification of virus like particles (VLPs) of foot-and-mouth disease virus in Eri silkworm (Samia cynthia ricini) larvae.
abstract: Foot-and-mouth disease (FMD) is a highly contagious viral disease, which causes severe economic loss to livestock. Virus like particles (VLPs) produced by recombinant DNA technology are gaining importance because of their immunogenic properties and safety in developing a new vaccine for FMD. In the present study, a practical and economically feasible approach of expression, purification and characterization of VLPs of FMDV in Eri silkworm (Samia cynthia ricini) larvae was described. Although three lepidopteran insect larvae (Helicoverpa armigera, Spodoptera litura and Samia cynthia ricini) were tested for production of VLPs, expression was obtained only in Eri silkworm larvae. High titred recombinant baculovirus encoding the polyprotein P1-2A-3C of FMDV was prepared in Sf9 cells. Injection of recombinant baculovirus into hemocoel of Eri silkworm larvae resulted in increasing levels of expression of VLPs in the hemolymph from 3 to 7 days post infection (dpi) compared to low level expression by oral feeding. The VLPs reacted in Sandwich ELISA with serum raised against whole virus particles of FMDV type O/IND/R2/75 and protein banding pattern of 26, 37 and 47 kDa in Western blotting demonstrated their antigenic resemblance to native virus. Sucrose density gradient purified VLPs were used for immunization of rabbits and guinea pigs for assessing immunogenicity. Further, the reactivity of serum samples of rabbits and guinea pigs in Indirect-ELISA with titres (1.30-2.81 Log10) indicated that the VLPs were antigenic and immunogenic in nature. We demonstrate that Eri silkworm larvae could be used for production of VLPs of FMDV type O/IND/R2/75 for the first time. This approach could be useful for large scale production of recombinant VLPs for vaccine or diagnostic use in FMD control programme.
authors.affiliation: Foot-and-Mouth Disease Vaccine Centre, Indian Veterinary Research Institute, Bangalore Campus, Hebbal, Bangalore, Karnataka 560024 India.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Foot-and-Mouth Disease Vaccine Centre, Indian Veterinary Research Institute, Bangalore Campus, Hebbal, Bangalore, Karnataka 560024 India.
authors.forename: P
authors.initials: P
authors.lastname: Saravanan
authors.affiliation: National Bureau of Agricultural Insect Resources, H A Farm Post, Bellary Road, Hebbal, Bangalore, Karnataka 560024 India.
authors.forename: S K
authors.initials: SK
authors.lastname: Jalali
keywords: Baculovirus expression; Eri silkworm larva; FMDV; Hemolymph; Purification of VLPs; Virus like particles (VLPs)