Sample Barcoding for Ultra High-Throughput Flow Cytometry

Project Summary High-throughput screens are the workhorse of the early drug discovery process. Millions of biochemical or cell-based assays are conducted to test the activity of small molecules, antibodies and oWher compoXndV Wo idenWif\ ³hiWV´ or drXg candidaWeV for fXrWher study, including pre-clinical and clinical studies. However, the most used technologies for high-throughput screening, such as HTRF, Alpha and ELISA, can only measure one or two parameters per sample, providing incompleWe informaWion of Whe drXg¶V biological mechanisms. Furthermore, these technologies detect analytes or activity outside the cell, which can result in misleading hits as compounds may fail to achieve the same effect in cell samples. To accelerate drug discovery, there is a major need for multiplexed screening at high throughput. The long-term goal of this application is to develop high-throughput flow cytometry to enable multiplexed drug screening. Although flow cytometry can measure up to 30 parameters per sample at single-cell resolution, its throughput has thus far been severely limited by its slow sampling speed. The short-term goal of Phase I is developing novel laser particle (LP) probes to barcode and pool cell samples to improve throughput, and demonstrate feasibility using a custom-built flow cytometer. This approach can eliminate sampling dead time, reducing acquisition time from 1 hour per 384-well plate to a few minutes. Furthermore, sample barcoding reduces reagent consumption and increases statistical robustness as samples are processed and stained together after pooling. Completion of Phase I research will establish the feasibility of high-throughput flow cytometry using LP barcodes. Development of this technology will greatly benefit the drug discovery industry by saving time and cost.Project Narrative Finding drug candidates commonly involves high-speed screening of millions of compounds on cell samples. Current screening technologies read only one or two parameters per sample, often insufficient to understand drug effects accurately. This project develops novel barcodes to enable more comprehensive screening and thereby contributing to faster drug discovery.