Recombinant protein expression in a Drosophila cell line: comparison with the baculovirus system.
In this report, we compare two different expression systems: baculovirus/Sf9 and stable recombinant Drosophila Schneider 2 (S2) cell lines. The construction of a recombinant S2 cell line is simple and quick, and in batch fermentations the cells have a doubling time of 20 hours until reaching a plateau density of 20 million cells/ml. Protein expression is driven by the Drosophila Metallothionein promoter which is tightly regulated. When expressed in S2 cells, the extracellular domain of human VCAM, an adhesion molecule, is indistinguishable from the same protein produced by baculovirus-infected Sf9 cells. Additionally, we present data on the expression of a seven trans-membrane protein, the dopamine D4 receptor, which has been successfully expressed in both systems. The receptor integrates correctly in the S2 membrane, binds [3H]spiperone with high affinity and exhibits pharmacological characteristics identical to that of the receptor expressed in Sf9 and mammalian cells. The general implications for large scale production of recombinant proteins are discussed.
Bernard
A R
AR
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
Kost
T A
TA
Overton
L
L
Cavegn
C
C
Young
J
J
Bertrand
M
M
Yahia-Cherif
Z
Z
Chabert
C
C
Mills
A
A
D000818:Animals; D016367:Baculoviridae; D015815:Cell Adhesion Molecules / Q000096:biosynthesis*; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D046508:Culture Techniques / Q000379:methods*; D004331:Drosophila melanogaster; D006801:Humans; D007700:Kinetics; D008668:Metallothionein / Q000235:genetics; D011401:Promoter Regions, Genetic; D011954:Receptors, Dopamine / Q000096:biosynthesis* / Q000378:metabolism; D017448:Receptors, Dopamine D2; D050638:Receptors, Dopamine D4; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000378:metabolism; D018411:Spodoptera; D014162:Transfection; D019010:Vascular Cell Adhesion Molecule-1
D015815:Cell Adhesion Molecules; C493599:DRD4 protein, human; D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2; D011994:Recombinant Proteins; D019010:Vascular Cell Adhesion Molecule-1; D050638:Receptors, Dopamine D4; D008668:Metallothionein