Early detection and metabolic pathway identification of T cell activation by in-process intracellular mass spectrometry

Austin L Culberson; Annie C Bowles-Welch; Bryan Wang; Peter A Kottke; Angela C Jimenez; Krishnendu Roy; Andrei G Fedorov

doi:10.1016/j.jcyt.2023.03.010

Early detection and metabolic pathway identification of T cell activation by in-process intracellular mass spectrometry

Cytotherapy. 2023 Sep;25(9):1006-1015. doi: 10.1016/j.jcyt.2023.03.010. Epub 2023 Apr 13.

Authors

Austin L Culberson¹, Annie C Bowles-Welch², Bryan Wang³, Peter A Kottke⁴, Angela C Jimenez⁵, Krishnendu Roy⁵, Andrei G Fedorov⁶

Affiliations

¹ The George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA; National Science Foundation Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA.
² Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA.
³ National Science Foundation Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA; Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA; The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA; School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
⁴ The George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA.
⁵ National Science Foundation Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA; Marcus Center for Therapeutic Cell Characterization and Manufacturing, Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia, USA; The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
⁶ The George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA; National Science Foundation Engineering Research Center (ERC) for Cell Manufacturing Technologies (CMaT), Atlanta, Georgia, USA. Electronic address: agf@gatech.edu.

PMID: 37061898
PMCID: PMC10524195 (available on 2024-09-01)
DOI: 10.1016/j.jcyt.2023.03.010

Abstract

Background aims: In-process monitoring and control of biomanufacturing workflows remains a significant challenge in the development, production, and application of cell therapies. New process analytical technologies must be developed to identify and control the critical process parameters that govern ex vivo cell growth and differentiation to ensure consistent and predictable safety, efficacy, and potency of clinical products.

Methods: This study demonstrates a new platform for at-line intracellular analysis of T-cells. Untargeted mass spectrometry analyses via the platform are correlated to conventional methods of T-cell assessment.

Results: Spectral markers and metabolic pathways correlated with T-cell activation and differentiation are detected at early time points via rapid, label-free metabolic measurements from a minimal number of cells as enabled by the platform. This is achieved while reducing the analytical time and resources as compared to conventional methods of T-cell assessment.

Conclusions: In addition to opportunities for fundamental insight into the dynamics of T-cell processes, this work highlights the potential of in-process monitoring and dynamic feedback control strategies via metabolic modulation to drive T-cell activation, proliferation, and differentiation throughout biomanufacturing.

Keywords: cell therapy; lab-on-a-chip; metabolic pathway; process analytical technology.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Cell Differentiation
Cell Proliferation
Mass Spectrometry
Metabolic Networks and Pathways*
T-Lymphocytes*

Grants and funding

R01 GM138802/GM/NIGMS NIH HHS/United States