High-level production of a functional immunoglobulin heterodimer in a baculovirus expression system.
A murine immunoglobulin heterodimer has been expressed in a baculovirus expression system. This was achieved by using both double infection of insect cells with separate heavy- and light-chain-expressing viruses and infection with a double-recombinant virus containing both the immunoglobulin heavy- and light-chain cDNAs. In both cases, the polypeptide chains were correctly processed, glycosylated, and assembled into normal H2L2 (H = heavy, L = light) immunoglobulin monomers. These molecules bound antigen and expressed both polyclonal idiotype and monoclonal idiotopes. Furthermore, the transfer vectors described have been modified to contain the F1 origin of replication for the production of single-stranded DNA, which facilitates site-specific mutations of either the polyhedrin promoter or the inserted foreign gene. Use of this system should significantly advance the analysis of the structural bases for both idiotype expression and antigen binding by immunoglobulin. More importantly, it provides a generic method for the high-level expression of antibodies of diverse interest.
Hasemann
C A
CA
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
Capra
J D
JD
D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007130:Immunoglobulin Idiotypes / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D009154:Mutation; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D011863:Radioimmunoassay; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
D007143:Immunoglobulin Heavy Chains; D007130:Immunoglobulin Idiotypes; D007147:Immunoglobulin Light Chains; D015345:Oligonucleotide Probes; D014764:Viral Proteins