A high density CHO-S transient transfection system: Comparison of ExpiCHO and Expi293.
Chinese Hamster Ovary (CHO) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. Until recently, production of milligrams to grams of protein in CHO transient systems was challenging. As such, Human Embryonic Kidney (HEK293) cells are the most common mammalian cell type used for transient transfection. The post-translational modifications (PTMs) of a protein are dictated in part by the cell line used for expression, and changes in PTMs have been shown to affect both the activity and biophysical properties of proteins. Therefore, it is potentially advantageous to keep the host cell type consistent throughout drug discovery and development. To this end, we compared the ExpiCHO system, a high density CHO-S transient transfection system, to the Expi293 and FreeStyle MAX CHO transient systems. Fourteen proteins were expressed in both the Expi293 and ExpiCHO systems. For a majority of proteins tested, the protein titers observed with the ExpiCHO system were higher than those seen with both the FreeStyle MAX CHO and Expi293 systems. Antibodies expressed using the ExpiCHO system had glycosylation patterns more similar to antibodies produced in stable CHO cell lines than Expi293-derived antibodies. However, culture duration and temperature were found to affect protein titer, monodispersity, enzyme activity, and PTMs and should be carefully selected when using the ExpiCHO system. The ExpiCHO transient transfection systems allows for facile production of milligrams to grams of protein in CHO cells and de-risks the transition from transient to stable material during drug development.
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Nina K
NK
Jain
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Susan
S
Barkowski-Clark
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Richard
R
Altman
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Krista
K
Johnson
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Fang
F
Sun
Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States.
Jonathan
J
Zmuda
Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States.
Chao Yan
CY
Liu
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Adriana
A
Kita
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Ryan
R
Schulz
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Alyssa
A
Neill
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Robert
R
Ballinger
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Rekha
R
Patel
Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States.
Jian
J
Liu
Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States.
Alinafe
A
Mpanda
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Brian
B
Huta
Cell Biology, Life Sciences Solutions Group, Thermo Fisher, 7335 Executive Way, Frederick, MD 21704, United States.
Henry
H
Chiou
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States.
Walter
W
Voegtli
Discovery Research CT, Alexion Pharmaceuticals, 100 College Street, New Haven, CT 06510, United States. Electronic address: Tadas.Panavas@alexion.com.
Tadas
T
Panavas
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins
CHO cells; Glycosylation; HEK293 cells; Post-translational modification; Transient transfection