Application note: Multiplexed detection of mouse cytokines using QBeads and the iQue Screener

This application note illustrates the value of multiplexing, rapid sampling, and assay miniaturisation using QBeads on the iQue Screener…

The use of rodent QBeads® Kits with the iQue® Screener enables multiplexed quantification of secreted mouse or rat cytokines in high throughput screening applications. Together, QBeads and the iQue Screener allow dramatic assay miniaturisation in microtiter plates, reducing reagent costs as well as minimising the required amount of expensive or difficult-to-obtain cell types. QBeads’ no-wash assay protocol combined with the iQue Screener’s rapid sampling technology allow more samples to be analysed in less time, making it practical to screen larger libraries, test more doses, and observe more time points.

Cells derived from rodents are important models in immunology research. In assays utilising these cells, profiling changes to cytokine levels are often key endpoints required to fully characterise an immune response. Many technologies exist to quantify cytokines, however, most lack features that are essential for high throughput screening. The use of QBeads on the iQue Screener allows multiplexed rodent cytokine quantification with three important benefits for screening: no-wash protocols, compatibility with a high degree of assay miniaturisation, and rapid plate sampling speeds.

QBeads are immunochemistry-based bead capture reagents that are able to quantify 28 different murine cytokines and 6 different rat cytokines. QBeads assays can be multiplexed to allow simultaneous quantification of multiple cytokines in a single assay, which preserves the ability to easily correlate results between different secreted proteins because a single assay setup is used for detection of all cytokines of interest. Using a single assay preparation for multiplexed detection also saves time and cost compared to running individual assays for each protein.

QBeads assays performed on the iQue Screener allow assay volumes to be drastically miniaturised. The iQue Screener’s proprietary rapid sampling technology allows assays performed in standard 384-well plates to be run at volumes around 9 µL. This level of assay miniaturisation is especially useful in high throughput screens that use primary rodent cells, where the total amount of cells harvested for the screen can limit the number of conditions that can be tested.