Separation of Activated T Cells Using Multidimensional Double Spiral (MDDS) Inertial Microfluidics for High-Efficiency CAR T Cell Manufacturing

Anal Chem. 2024 Jul 2;96(26):10780-10790. doi: 10.1021/acs.analchem.4c01981. Epub 2024 Jun 18.

Abstract

This study introduces a T cell enrichment process, capitalizing on the size differences between activated and unactivated T cells to facilitate the isolation of activated, transducible T cells. By employing multidimensional double spiral (MDDS) inertial sorting, our approach aims to remove unactivated or not fully activated T cells post-activation, consequently enhancing the efficiency of chimeric antigen receptor (CAR) T cell manufacturing. Our findings reveal that incorporating a simple, label-free, and continuous MDDS sorting step yields a purer T cell population, exhibiting significantly enhanced viability and CAR-transducibility (with up to 85% removal of unactivated T cells and approximately 80% recovery of activated T cells); we found approximately 2-fold increase in CAR transduction efficiency for a specific sample, escalating from ∼10% to ∼20%, but this efficiency highly depends on the original T cell sample as MDDS sorting would be more effective for samples possessing a higher proportion of unactivated T cells. This new cell separation process could augment the efficiency, yield, and cost-effectiveness of CAR T cell manufacturing, potentially broadening the accessibility of this transformative therapy and contributing to improved patient outcomes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Separation* / methods
  • Humans
  • Immunotherapy, Adoptive / methods
  • Lymphocyte Activation*
  • Microfluidic Analytical Techniques / instrumentation
  • Receptors, Chimeric Antigen* / metabolism
  • T-Lymphocytes* / cytology

Substances

  • Receptors, Chimeric Antigen