Effects of co-expressing chaperone BiP on functional antibody production in the baculovirus system.
The assembly pathway of the insect cell Spodoptera frugiperda (Sf-9) was engineered to include expression of the murine chaperone immunoglobulin heavy chain binding protein (BiP) using the baculovirus vector. The impact of BiP coexpression on the production and secretion of functional and soluble recombinant immunoglobulin IgG levels was evaluated. Recombinant BiP was found to associate specifically with immunoglobulins in immunoprecipitation studies. Coinfection of insect cells with a BiP-containing baculovirus and baculoviruses coding for two different murine IgG proteins increased intracellular functional antibody activity levels substantially above the levels observed in the absence of BiP. Soluble intracellular immunoglobulin levels were found to increase as well. However, secreted functional antibody levels did not increase significantly. Also, degradation of heavy chain immunoglobulin in insect cells was indicated by the accumulation of lower molecular weight immunoglobulins at 4 days postinfection. Coexpression of light chains reduced the level of these lower molecular weight immunoglobulins while BiP coexpression led to enhanced levels. These findings suggest that coexpressed BiP can increase intracellular soluble and functional antibody yields but that secretion in the baculovirus-insect cell system must be limited at some post-translational step.
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218-2694.
T A
TA
Hsu
J J
JJ
Eiden
P
P
Bourgarel
T
T
Meo
M J
MJ
Betenbaugh
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D007118:Immunoassay; D007136:Immunoglobulins / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D011995:Recombination, Genetic; D018411:Spodoptera
D007136:Immunoglobulins; D018832:Molecular Chaperones