Complementation of recombinant baculoviruses by coinfection with wild-type virus facilitates production in insect larvae of antigenic proteins of hepatitis B virus and influenza virus.
We describe the coinfection of insects with wild-type and recombinant baculoviruses in which the polyhedrin gene promoter is used to express hepatitis B virus envelope protein (hepatitis B virus surface antigen; HBsAg) or influenza A virus neuraminidase (NA). Viruses were administered per os to larvae of the cabbage looper, Trichoplusia ni, causing an infection that within 5 days resulted in the production of approximately 0.15 mg of HBsAg per insect, representing 1.5% of the total extracted protein, or approximately 2.8 mg of NA per insect, representing 28% of the total extractable protein. The HBsAg and NA produced by infected larvae were purified from insect lysates. These proteins were antigenic as determined by conformation-dependent immunoassays. The NA was enzymatically active with conventional substrates. The method of infection described allows genetic complementation by wild-type virus of recombinant viruses lacking the polyhedrin gene essential for infection per os and has implications for the high-yield production in insect larvae of other recombinant proteins of baculoviruses.
Price
P M
PM
Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029.
Reichelderfer
C F
CF
Johansson
B E
BE
Kilbourne
E D
ED
Acs
G
G
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics*; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D007700:Kinetics; D007814:Larva; D009439:Neuraminidase / Q000235:genetics* / Q000378:metabolism
D006514:Hepatitis B Surface Antigens; D009439:Neuraminidase