Antibody Production in Silkworm Cells and Silkworm Larvae Infected with a Dual Recombinant Bombyx Mori Nuclear Polyhedrosis Virus

Abstract

We have examined the efficiency of coexpression of two heterologous genes from a recombinant Bombyx mori nuclear polyhedrosis virus for the production of antibodies in silkworm larvae. The cDNAs encoding the light and the heavy chains of a murine immunoglobulin, directed against lipoprotein I of Pseudomonas aeruginosa, were brought under the control of two separate copies of the viral polyhedrin promotor. Infection of silkworm cells with the recombinant baculovirus yielded a maximum of 6.4 micrograms/ml IgG2A in the culture supernatant 72 hours post infection, while 800 micrograms/ml IgG2A was found in the hemolymph of infected fifth instar silkworm larvae seven days after infection with the same construct. The recombinant antibody exhibited a similar antigen specificity and avidity to that of the monoclonal antibody derived from ascites fluid.

In VitroandIn VivoSecretion of Cloned Antibodies by Genetically Modified Myogenic Cells

“…Although production of antibodies has been documented in the case of a few cell Unes such as COS-7 ceUs (see Noel et al, 1996, for example) and nonmammaUan cells (Ma et al, 1995;Reis et al, 1992;Nesbit et al, 1992), tiiis is to our knowledge the first time that cells suitable for gene therapy protocols have been utiUzed, i.e., cells (i) that can be withdrawn from patients, (ii) that are amenable to stable gene ttansfer, (iu) that can be used for autologous grafts, and (iv) that can persist on the long-term in grafted patients. When implanted in the foreUmb of a mouse, engineered myogenic cells, including primary myoblasts, were also shown capable of deUvering antibodies into the bloodstteam.…”

Bombyx mori Nucleopolyhedrovirus Bacmid Enabling Rapid Generation of Recombinant Virus by In Vitro Transposition

“…This could have benefits for the production of functional or authentic proteins [8]. Second, the yields obtained in insect larvae could be up to 500-fold higher than those obtained in cultured cells [4,11,16]. Finally, expression using insect larvae represents a relatively low cost alternative to the production of a foreign protein using cultured cells [4,8].…”

Production of Antibody in Insect Cells

“…Because a recombinant baculovirus can also infect insect larvae or pupae, they have been used for recombinant protein production as "protein factories" (Kato et al, 2010;Maeda et al, 1985;Medin et al, 1990). Upon infection of silkworm larvae using a needle moistened with a recombinant BmNPV, a functional IgG has been expressed at a concentration of 800 g/ml in the hemolymph (Reis et al, 1992) (Table 2). A functionally active Fab fragment has been produced, with a yield of 1.1 mg/g-larvae, upon infection of T. ni larvae after the ingestion of a diet containing a recombinant AcNPV (O'Connell et al, 2007).…”