Two-color selection for amplified co-production of proteins in mammalian cells.
Structural and functional analyses for many mammalian systems depend on having abundant supplies of recombinant multi-protein complexes that can be produced best, or only, in mammalian cells. We present an efficient fluorescence marking procedure for establishing stable cell lines that overexpress two proteins in co-ordination, and we validate the method in the production of monoclonal antibody Fab fragments. The procedure has worked without fail on all seven of seven trials on Fabs, which are being used in the crystallization of G-protein coupled receptors. This manner of efficient selection may readily be adapted for the co-production of other complexes of two or more proteins.
Assur
Zahra
Z
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032, USA.
Schieren
Ira
I
Hendrickson
Wayne A
WA
Mancia
Filippo
F
D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002460:Cell Line; D002469:Cell Separation; D003116:Color; D005453:Fluorescence; D006801:Humans; D018613:Microscopy, Confocal; D011506:Proteins / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011506:Proteins