Inertial Microfluidic Purification of CAR-T-Cell Products

Mona T Elsemary; Michelle F Maritz; Louise E Smith; Majid Warkiani; Veronika Bandara; Silvana Napoli; Simon C Barry; Justin T Coombs; Benjamin Thierry

doi:10.1002/adbi.202101018

Inertial Microfluidic Purification of CAR-T-Cell Products

Adv Biol (Weinh). 2022 Jan;6(1):e2101018. doi: 10.1002/adbi.202101018. Epub 2021 Dec 8.

Authors

Mona T Elsemary¹, Michelle F Maritz², Louise E Smith³, Majid Warkiani⁴, Veronika Bandara⁵, Silvana Napoli⁵, Simon C Barry⁵, Justin T Coombs⁶, Benjamin Thierry²

Affiliations

¹ Future Industries Institute, ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Cell Therapy Manufacturing Cooperative Research Centre, University of South Australia Mawson Lakes Campus, Mawson Lakes, SA, 5095, Australia.
² Future Industries Institute, ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of South Australia Mawson Lakes Campus, Mawson Lakes, SA, 5095, Australia.
³ Future Industries Institute, Cell Therapy Manufacturing Cooperative Research Centre, University of South Australia Mawson Lakes Campus, Mawson Lakes, SA, 5095, Australia.
⁴ School of Biomedical Engineering, University of Technology Sydney, Broadway, Ultimo, NSW, 2007, Australia.
⁵ Women's and Children's Hospital, Adelaide, SA, 5006, Australia.
⁶ Carina Biotech, Adelaide, SA, 5095, Australia.

PMID: 34881810
DOI: 10.1002/adbi.202101018

Abstract

Chimeric antigen receptor T (CAR-T) cell therapy is rapidly becoming a frontline cancer therapy. However, the manufacturing process is time-, labor- and cost-intensive, and it suffers from significant bottlenecks. Many CAR-T products fail to reach the viability release criteria set by regulators for commercial cell therapy products. This results in non-recoupable costs for the manufacturer and is detrimental to patients who may not receive their scheduled treatment or receive out-of-specification suboptimal formulation. It is demonstrated here that inertial microfluidics can, within minutes, efficiently deplete nonviable cells from low-viability CAR-T cell products. The percentage of viable cells increases from 40% (SD ± 0.12) to 71% (SD ± 0.09) for untransduced T cells and from 51% (SD ± 0.12) to 71% (SD ± 0.09) for CAR-T cells, which meets the clinical trials' release parameters. In addition, the processing of CAR-T cells formulated in CryStor yields a 91% reduction in the amount of the cryoprotectant dimethyl sulfoxide. Inertial microfluidic processing has no detrimental effects on the proliferation and cytotoxicity of CAR-T cells. Interestingly, ≈50% of T-regulatory and T-suppressor cells are depleted, suggesting the potential for inertial microfluidic processing to tune the phenotypical composition of T-cell products.

Keywords: CAR-T cell; cryoprotectant; manufacture; microfluidic; purification; viability.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell- and Tissue-Based Therapy
Humans
Immunotherapy, Adoptive
Lymphocyte Count
Microfluidics
Receptors, Chimeric Antigen*

Substances

Receptors, Chimeric Antigen