Multiplexed Cell- and Bead-based Assays in a No-wash Format for Biologics Screening and Characterization
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, IL-6 and IL-8 cytokine detection assay where reagents from two commercially available single-plex ELISA kits were transferred onto TTP Labtech sol-R™ coded beads and multiplexed together. The results of this alternative fluorescence-based approach demonstrate similar performance to ELISA, but with a significantly streamlined work-flow.
Overall, the data presented here show the versatile nature of and advantages of multiplexing assays using TTP Labtech’s mirrorball for improving the productivity of biologics discovery.
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, IL-6 and IL-8 cytokine detection assay where reagents from two commercially available single-plex ELISA kits were transferred onto TTP Labtech sol-R™ coded beads and multiplexed together. The results of this alternative fluorescence-based approach demonstrate similar performance to ELISA, but with a significantly streamlined work-flow.
Overall, the data presented here show the versatile nature of and advantages of multiplexing assays using TTP Labtech’s mirrorball for improving the productivity of biologics discovery.
