# Two-stage inertial microfluidics enrichment of activated T-cells towards a bead-less chimeric antigen receptor manufacturing protocol

## TL;DR

This paper introduces a new bead-free method using microfluidics to enrich T-cells for CAR-T therapy, which could simplify and improve the manufacturing process.

## Contribution

A novel two-stage inertial microfluidics method for T-cell enrichment without using beads, offering high purity and recovery.

## Key findings

- The dual-stage process achieved 87% T-cell enrichment and 80% recovery from healthy donor samples.
- Validation with ovarian cancer samples showed 70% T-cell purity improvement with 64% recovery.
- The method had no detectable effect on T-cell proliferation.

## Abstract

CAR-T cell therapy is leading the way in the field of cancer cell immunotherapies due to its high success rates. However, the manufacturing of CAR-T cells remains complex and expensive. T-cell enrichment from patient apheresis starting material is a key step in the manufacture but cellular impurities interfere with the ex vivo transduction of T-cells and their proliferation. Current enrichment methods including magnetic bead selection suffer from various limitations. We report here a bead-less T-cell enrichment process through a two-stage procedure based on inertial microfluidics. Using apheresis like starting material samples from healthy donors, the dual-stage process showed an efficient 87% (SD ± 6%) enrichment and 80% (SD ± 30%) recovery of T-cells. Validation of the process with ovarian cancer samples resulted in a T-cell purity 70% (SD ± 10%) from a starting purity of 48% (SD ± 6%) at a 64% (SD ± 4%) T-cell recovery. The two-stage inertial microfluidic process was also shown to have no detectable effect on the proliferation of the cells.