mirrorball: simultaneous determination of antibody binding, specificity and titer on the mirrorball fluorescence cytometer

Biologics such as antibodies are ideally suited for therapeutic use against cell-surface disease targets. When screening, the use of cells expressing native target proteins minimises the risk of false positive binding (which can occur when using recombinant proteins) leading to improved hit quality.

Traditional screening methods to identify and characterise hits often report a single measurement per well, due to either assay or instrumentation constraints. Multiplexed assay formats facilitate the reporting of multiple readouts to further improve data quality and enhance productivity, whilst also conserving sample.

Here we present the results generated from multiplexed no-wash protocols using TTP Labtech’s mirrorball®, a laser scanning cytometer. To model antibody screening, anti-EGFR antibodies were titrated against multiplexed target-expressing and control cells to determine specific versus non-specific binding in one productive screen. Additionally, we report the results of a no-wash, cytokine detection assay where reagents from two commercially available singleplex ELISA kits were transferred onto TTP Labtech sol-R™ coded beads and multiplexed.
Results demonstrate similar performance to ELISA, but with a streamlined workflow process.

Overall, the data presented here show the versatile nature, and advantages of multiplexing assays using TTP Labtech’s mirrorball for improving the productivity of biologics discovery.