I-Corps: A label-free, autofluorescence lifetime flow cytometer for in-line monitoring of T cell function

University Of Wisconsin-Madison, Madison WI

Investigators

Abstract

The broader impact/commercial potential of this I-Corps project is the development of a label-free method for cell therapy manufacturing process optimization and product characterization prior to clinical use. Cell-based therapies have the potential to treat, or even cure, a myriad of diseases. However, these complex biological products display intrinsic variability within a tightly regulated industry and standard quality assessments require labels to characterize functional subsets of cells. Current methods of assessment are laborious, time-consuming, susceptible to reagent quality variability, and may alter cell function. The proposed technology may provide a solution for cell manufacturing that is critical for the continued development of cell-based therapies. This I-Corps project is based on the development of a label-free, non-destructive optical detection technology with single-cell resolution to quantify cell function, viability, and metabolism including machine learning methods that predict later cell quality. The proposed hardware, optics, and analytical algorithms may be readily integrated into a variety of geometries, such as flow cytometry and microscopy, enabling non-destructive assessment for rapid, label-free diagnostics and biomanufacturing process optimization. The inherent flexibility of the proposed technology enables non-destructive assessment for rapid, label-free biomanufacturing process optimization. In addition, a prototype has been engineered that was purpose-built to integrate with biomanufacturing workflows at reduced cost and footprint compared to commercial instruments. Software also has been developed to detect functional biomarker changes at the cell level along with machine learning models that predict later cell function, neither of which are commercially available. The proposed technology operates on intrinsic universal cellular signals, enabling utility across all cell types. Previous results demonstrated that the technology is sensitive to functional response from T cells that predict activation within minutes, preceding expression of activation surface marker at 1 hour. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.