Functional single-cell screening of chimeric antigen receptor (CAR)-T cells based on multiple cytokines using Nanovial Technology

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B928
Abstract ID: 5662

Presenting Author:
Wei-Ying Kuo , Senior Scientist at Partillion Bioscience Corp.

Abstract:

CAR-T therapies, while potent against some cancers, face challenges including toxicities and tumor relapse, particularly in solid tumors with hostile TMEs and sub-optimal CAR designs. A major obstacle is the lack of efficient functional screening methods for CAR-T cells, crucial for T-cell-based immunotherapy progress.

Our study introduces a novel functional screening approach using hydrogel microparticles (Nanovials) to analyze millions of individual CAR-T cells based on antigen specificity and functional cytokine secretion. We modified Nanovials with Mesothelin (MSLN) antigen, enriching and binding specific CAR-T cells. Additionally, Nanovials were adapted with antibodies to detect key cytokines (IFN-gamma, GranzymeB) from activated CAR-T cells, identified through fluorescent markers. This setup allows for analyzing and sorting CAR-T cells with unique functional traits using standard flow cytometers.

Using the Nanovial assay we distinguished MSLN CAR-T variants by cytokine profiles, revealing minor CAR engineering differences and enabling early functional assessment. This technique identified distinct CAR-T cell subpopulations within patient samples, surpassing traditional bulk analysis methods like ELISA and cell killing assays.

In summary, our Nanovial-based method offers a scalable, efficient solution for CAR-T cell functional screening, enhancing CAR design and improving therapeutic manufacturing quality control.