Localized Sampling Enables Monitoring of Cell State via Inline Electrospray Ionization Mass Spectrometry

• Type of Material: Article
• Authors: Mason A Chilmonczyk, Georgia Institute of Technology; Gilad Doron, Georgia Institute of Technology; Peter A Kottke, Georgia Institute of Technology; Austin L Culberson, Georgia Institute of Technology; Kelly Leguineche, University of Oregon; Robert Guldberg, Emory University; Edwin Horwitz, Emory University; Andrei G Fedorov, Georgia Institute of Technology
• Language: English
• Date: 2020-10-12
• Publisher: WILEY-V C H VERLAG GMBH
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2020 Wiley‐VCH GmbH
• Final Published Version (URL): http://dx.doi.org/10.1002/biot.202000277
• Title of Journal or Parent Work: BIOTECHNOLOGY JOURNAL
• Volume: 16
• Issue: 3
• Start Page: e2000277
• End Page: e2000277
• Grant/Funding Information: The work described is supported by NSF Center for Cell Manufacturing Technologies (CMaT) Award 1648035, Marcus Center for Therapeutic Cell Characterization and Manufacturing Collaboration Grant in Cell Manufacturing, The Georgia Tech Foundation, and the Georgia Research Alliance. Partial support was also provided by Grant Number RO1GM112662 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH. Device micro-fabrication was performed in part at the Georgia Tech Institute for Electronics and Nanotechnology, a member of the National Nanotechnology Coordinated Infrastructure, which is supported by the National Science Foundation (Grant ECCS-1542174).
• Keywords: online electrospray ionization mass spectrometry; SECRETOME; MINERALIZATION; Biochemistry & Molecular Biology; cell manufacturing; Biochemical Research Methods; PROTEOMICS; MESENCHYMAL STEM-CELLS; in situ secretome analysis; cell state and quality attributes; Science & Technology; Biotechnology & Applied Microbiology; DIFFERENTIATION; Life Sciences & Biomedicine; real-time biomarker monitoring and discovery

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.