Production of recombinant proteins by baculovirus-infected gypsy moth cells.
An experimental study was undertaken to evaluate alternative insect cell lines to Sf9 [from Spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. Insect cell lines from two different organisms were considered: IPLB-LdEIta (LdEIta) from Lymantria dispar (gypsy moth) and IPLB-HvT1 (HvT1) from Heliothis virescens (tobacco budworm). Both LdEIta and HvT1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than Sf9 after infection with the Autographa californica nuclear polyhedrosis virus (AcMNPV). However, only LdEIta generated a product yield (activity per milligram of total protein) which exceeded that of Sf9 (by 25%), so its growth and production characteristics were investigated in depth. LdEIta generated production levels and yields of a recombinant rotaviral protein, VP4, which exceeded those of Sf9 by 84 and 38%, respectively. In suspension culture, the LdEIta cells grew as aggregates with a doubling time several hours longer than Sf9, but the recombinant product yields of LdEIta were still higher than Sf9 by 38% in this culture environment. beta-Galactosidase expression rates and cell death rates suggested that the difference in productivity between the two hosts was due to the ability of LdEIta to survive the baculovirus infection and produce recombinant proteins longer than Sf9. The presence of LdEIta aggregates in suspension culture may be used as a method to separate live cells from dead cells, labile product, and spent medium in recombinant protein production processes.
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
M J
MJ
Betenbaugh
L
L
Balog
P S
PS
Lee
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002449:Cell Aggregation; D002460:Cell Line; D005822:Genetic Vectors; D009036:Moths / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D001616:beta-Galactosidase