Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines.
Recombinant Autographa california baculoviruses expressing genes for pseudorabies virus glycoprotein (gp50T), human plasminogen (HPg), and beta-galactosidase (beta-gal) were used to infect 23 cell lines or strains. The objectives were to compare amounts of recombinant proteins expressed in the cell lines, compare yields from clones and parent lines, investigate the effects of long-term culture in serum-free medium on production, and determine if some lines yield gp50T with different glycosylation patterns. For HPg, IZD-MB0503 had the highest yield and four other lines (IPLB-TN-R2, IPLB-SF-1254, IPLB-LdEIta, and CM-1) had levels above that of SF-9 cells. For gp50T, four lines (IPLB-HvT1, IPLB-SF21AE, IPLB-SF21AE-15, and IPLB-SF-1254) had higher amounts than SF-9 cells. Some lines yielded gp50T with molecular mass about 1000 daltons larger than that from SF-9 cells, which suggests increased oligosaccharide processing. Equally high levels of beta-gal were expressed in three lines (SF-9, IZD-MB0503, and BCIRL-PX2-HNV3). The major conclusion is that no single cell line produced highest yields for all three recombinant proteins. Four lines were cultured in serum-free medium for 31-34 passages and then infected with the three recombinant viruses. For most cell line-recombinant combinations, the yields in serum-free medium were equal to or better than those in serum-supplemented medium. Medium composition had a much stronger effect on foreign gene expression than on susceptibility of cells to wild-type virus.
Hink
W F
WF
Department of Entomology, Ohio State University, Columbus 43210.
Thomsen
D R
DR
Davidson
D J
DJ
Meyer
A L
AL
Castellino
F J
FJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001769:Blood; D002460:Cell Line; D002999:Clone Cells; D003470:Culture Media; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D007313:Insecta / Q000378:metabolism*; D010958:Plasminogen / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D003470:Culture Media; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C042966:pseudorabies virus glycoproteins; D010958:Plasminogen; D001616:beta-Galactosidase