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Localized Sampling Enables Monitoring of Cell State via Inline Electrospray Ionization Mass Spectrometry |
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| Authors: | Mason A Chilmonczyk Gilad Doron Peter A Kottke Austin L Culberson Kelly Leguineche Robert E Guldberg Edwin M Horwitz Andrei G Fedorov |
| Institution: | 1. The George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332 USA;2. NSF Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA, 30332 USA;3. The George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332 USA
NSF Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA, 30332 USA;4. The Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, 97403 USA;5. NSF Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA, 30332 USA The Phil and Penny Knight Campus for Accelerating Scientific Impact, University of Oregon, Eugene, OR, 97403 USA;6. Department of Pediatrics, Emory University, Atlanta, GA, 30332 USA |
| Abstract: | Nascent advanced therapies, including regenerative medicine and cell and gene therapies, rely on the production of cells in bioreactors that are highly heterogeneous in both space and time. Unfortunately, advanced therapies have failed to reach a wide patient population due to unreliable manufacturing processes that result in batch variability and cost prohibitive production. This can be attributed largely to a void in existing process analytical technologies (PATs) capable of characterizing the secreted critical quality attribute (CQA) biomolecules that correlate with the final product quality. The Dynamic Sampling Platform (DSP) is a PAT for cell bioreactor monitoring that can be coupled to a suite of sensor techniques to provide real-time feedback on spatial and temporal CQA content in situ. In this study, DSP is coupled with electrospray ionization mass spectrometry and direct-from-culture sampling to obtain measures of CQA content in bulk media and the cell microenvironment throughout the entire cell culture process (≈3 weeks). Post hoc analysis of this real-time data reveals that sampling from the microenvironment enables cell state monitoring (e.g., confluence, differentiation). These results demonstrate that an effective PAT should incorporate both spatial and temporal resolution to serve as an effective input for feedback control in biomanufacturing. |
| Keywords: | cell manufacturing cell state and quality attributes in situ secretome analysis online electrospray ionization mass spectrometry real-time biomarker monitoring and discovery |