Exploring the linkage between cell culture process parameters and downstream processing utilizing a plackett-burman design for a model monoclonal antibody.
Linkage of upstream cell culture with downstream processing and purification is an aspect of Quality by Design crucial for efficient and consistent production of high quality biopharmaceutical proteins. In a previous Plackett-Burman screening study of parallel bioreactor cultures we evaluated main effects of 11 process variables, such as agitation, sparge rate, feeding regimens, dissolved oxygen set point, inoculation density, supplement addition, temperature, and pH shifts. In this follow-up study, we observed linkages between cell culture process parameters and downstream capture chromatography performance and subsequent antibody attributes. In depth analysis of the capture chromatography purification of harvested cell culture fluid yielded significant effects of upstream process parameters on host cell protein abundance and behavior. A variety of methods were used to characterize the antibody both after purification and buffer formulation. This analysis provided insight in to the significant impacts of upstream process parameters on aggregate formation, impurities, and protein structure. This report highlights the utility of linkage studies in identifying how changes in upstream parameters can impact downstream critical quality attributes. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:163-170, 2017.
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Cyrus D
CD
Agarabi
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Brittany K
0000-0002-4233-1946
BK
Chavez
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Scott C
SC
Lute
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Erik K
EK
Read
Div. of Pharmaceutical Analysis, Office of Testing and Research, OPQ, CDER, FDA, Silver Spring, MD.
Sarah
S
Rogstad
Div. of Post Marketing Activities II, OPQ, CDER, FDA, Silver Spring, MD.
David
D
Awotwe-Otoo
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Matthew R
MR
Brown
Div. of Pharmaceutical Analysis, Office of Testing and Research, OPQ, CDER, FDA, Silver Spring, MD.
Michael T
MT
Boyne
Div. II, Office of Biotechnology Products, Office of Pharmaceutical Quality, CDER, FDA, Silver Spring, MD.
Kurt A
KA
Brorson
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D003412:Cricetulus; D006863:Hydrogen-Ion Concentration; D013696:Temperature
D000911:Antibodies, Monoclonal
design of experiments (DoE); host cell protein (HCP); mammalian cell culture; monoclonal antibody; quality by design (QbD)