Higher expression of Fab antibody fragments in a CHO cell line at reduced temperature.
A chimeric Fab was expressed in Chinese hamster ovary cells under the control of the CMV promoter in a two-stage production process. Cells were first grown to 90% confluence at 37 degrees C in a proliferation phase, followed by a production phase at either 37 degrees C or 28 degrees C. Medium supplemented with serum and medium free from serum was tested in the production phase at both temperatures. Comparison of Fab expression revealed that reducing the temperature to 28 degrees C resulted in a 14-fold increase in product yield when cells were cultivated in serum-containing medium, and in a 38-fold increase in product yield when serum-free medium was applied.
Schatz
Simone M
SM
Baxter BioScience, Biomedical Research Center, Orth/Donau, Austria.
Kerschbaumer
Randolf J
RJ
Gerstenbauer
Gabriele
G
Kral
Martina
M
Dorner
Friedrich
F
Scheiflinger
Friedrich
F
D000818:Animals; D019149:Bioreactors / Q000382:microbiology*; D016466:CHO Cells / Q000166:cytology* / Q000378:metabolism*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013696:Temperature
D016895:Culture Media, Serum-Free; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins