True screening for immunology: transforming science with a practical tool for large-scale discovery

Enormous effort is being put into understanding the interactions that make up the complex communications of the immune system so that they can be used as targets for therapeutic intervention. Whether the goal is finding a new therapeutic antibody, developing a specific checkpoint inhibitor, or evaluating which modified CAR – T cell population is killing a patient’s cancer cells, the need to evaluate more samples in a biologically relevant, reproducible and cost effective way has become acute.

Flow cytometry, the gold standard of detection in immunology research for decades, provides a high degree of data content on cell phenotypes and functions, and yet suffers limitations in terms of assay throughput, automation, sample/reagent volume requirements, and ease of use. Even advances in microplate auto-sampling for standard flow fails to address issues of reducing reagent cost, expertise required for assay setup, and dealing effectively with plate based data analysis and visualisation of results. Because these issues remain, the broad adoption of flow cytometry for screening applications has been hampered.

IntelliCyt Corporation has developed a high throughput/content screening platform for suspension cell and bead assays. The iQue Screener PLUS platform combines a novel, patented rapid micro-volume sampling technology, an optimised flow based ‘detection engine’ (three lasers, 15 parameters, no PMT adjustments), and a microplate centric assay development, analysis, and visualisation work environment. True immunology screening at high throughput levels (384 well plates processed in as little as 15 minutes) with reagent saving assay volumes (as little as six to nine µL/well) is routinely being undertaken in all of the top ten pharmaceutical companies with IntelliCyt technology. ForeCyt software provides plate level annotation allowing normalisations to controls, standard curve based quantitation and IC/EC50 curve generation. Assay development is carried out on an entire plate of data and any gate adjustments update results in real time, not one well at a time. A unique, high content data visualisation tool allows multiple data endpoints to be queried with boolean logic against user defined criteria, instantly locating active wells. All visualisations can be dynamically linked for easy biological confirmation.

Three main application areas for immunology screening are antibody discovery, immune targets, and immunotherapy. In the antibody discovery area, IntelliCyt technology is used to increase the throughput and content of antibody library screens, derived from multiple sources (hybridoma, phage display). By using antigens presented on cell surfaces and multiplexing cell populations with various controls in each well, antibody specificity and species cross reactivity can be assessed simultaneously, dramatically increasing data quality while reducing the time to screen a library from weeks to days. In the immune targets area, users have been able to reduce reagent consumption for cytokine profiling and combine cytokine assessment with functional assays on phenotyped cell subpopulations in the same wells. These assays have been formulated as no wash assays and can be miniaturised, saving money and time. In the immunotherapy area, profiling of modified CAR-T cells and TIL’s to kill cancer cells can take place rapidly, with a high degree of multiplexing, enabling better decisions on which populations to expand for therapeutic return to the patient. IntelliCyt is making a difference!