Transient transfection of CHO-K1-S using serum-free medium in suspension: a rapid mammalian protein expression system.
With the recent completion of the human genome sequencing project, scientists now face the daunting challenge of deciphering the function of these newly found genes quickly and efficiently. For biotechnology, it is equally important to identify the therapeutically relevant genes as quickly as possible. Mammalian expression systems provide many advantages to aid in this task. Mammalian cell lines have the capacity for proper post-translational modifications, including proper protein folding and glycosylation. In response to these needs, a CHO-K1 cell line that grows in suspension and in serum-free media was initially established and designated CHO-K1-S. An antibody gene of interest was chosen as the target for optimization rather than a reporter gene system. A comparison of various lipid transfection reagents was made using recombinant protein expression as the endpoint readout. Various other parameters including lipid:DNA ratios, cell density, and transfections in shaker versus spinner flasks were tested using the CHO-K1-S cell line. As a result, a rapid and reliable transient transfection protocol was developed. Using this procedure, we have produced milligram/per liter quantities of bioactive recombinant proteins from several genes of interest.
Rosser
Mary P
MP
Cell Sciences Department, Berlex Biosciences, Richmond, CA 94804, USA.
Xia
Wei
W
Hartsell
Steven
S
McCaman
Michael
M
Zhu
Ying
Y
Wang
Soujuan
S
Harvey
Susan
S
Bringmann
Peter
P
Cobb
Ronald R
RR
D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007202:Indicators and Reagents; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics*; D014162:Transfection / Q000295:instrumentation / Q000379:methods*
D016895:Culture Media, Serum-Free; D007141:Immunoglobulin Fc Fragments; D007202:Indicators and Reagents; D011994:Recombinant Proteins