title: High-Throughput, Automated Protein A Purification Platform with Multiattribute LC-MS Analysis for Advanced Cell Culture Process Monitoring.
abstract: The levels of many product related variants observed during the production of monoclonal antibodies are dependent on control of the manufacturing process, especially the cell culture process. However, it is difficult to characterize samples pulled from the bioreactor due to the low levels of product during the early stages of the process and the high levels of interfering reagents. Furthermore, analytical results are often not available for several days, which slows the process development cycle and prevents "real time" adjustments to the manufacturing process. To reduce the delay and enhance our ability to achieve quality targets, we have developed a low-volume, high-throughput, and high-content analytical platform for at-line product quality analysis. This workflow includes an automated, 96-well plate protein A purification step to isolate antibody product from the cell culture fermentation broth, followed by rapid, multiattribute LC-MS analysis. We have demonstrated quantitative correlations between particular process parameters with the levels of glycosylated and glycated species in a series of small scale experiments, but the platform could be used to monitor other attributes and applied across the biopharmaceutical industry.
authors.affiliation: Large Molecule Analytical Development, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Malvern, Pennsylvania 19355, United States.
authors.forename: Jia
authors.initials: J
authors.lastname: Dong
authors.affiliation: Active Pharmaceutical Ingredient Large Molecule, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Malvern, Pennsylvania 19355, United States.
authors.forename: Nicole
authors.initials: N
authors.lastname: Migliore
authors.affiliation: Strategic Operations, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Spring House, Pennsylvania 19477, United States.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Mehrman
authors.affiliation: Material Science, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Malvern, Pennsylvania 19355, United States.
authors.forename: John
authors.initials: J
authors.lastname: Cunningham
authors.affiliation: Large Molecule Analytical Development, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Malvern, Pennsylvania 19355, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Lewis
authors.affiliation: Large Molecule Analytical Development, Pharmaceutical Development & Manufacturing Science, Janssen Research & Development , Malvern, Pennsylvania 19355, United States.
authors.forename: Ping
authors.initials: P
authors.lastname: Hu
mesh_terms: D000818:Animals; D001331:Automation; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D003412:Cricetulus; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D013058:Mass Spectrometry / Q000295:instrumentation; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D013205:Staphylococcal Protein A