Quantification of biologically relevant secretable proteins and cytokines is critical in basic research and drug development, including understanding T-cell activation and cellular signaling cascades in inflammation and oncology. Over the last decade there has been an enormous expansion of research focused on inflammatory process and diseases conditions in which cytokines are measured and analyzed. These studies revolve around the ability to detect, quantify, and discriminate specific or multiple cytokines from a multitude of biomolecules present in any given sample. Researchers currently employ a variety of established immunoassay techniques such as Enzyme-Linked Immunosorbent Assay (ELISA), multiplex and flow cytometry-based assays to analyze the level of secreted protein in various disease models and exploratory research areas, however, traditional techniques for measuring protein secretion:
- Are low-throughput, laborious and time-consuming, requiring steps such as labelling, protocol optimization, long incubations and repetitive washes
- Require centrifugation steps to separate supernatants from cells
- Are generally limited to analyzing one analyte per sample, increasing data variability and necessitating the use of large volumes of precious sample and reagents
- Produce data requiring complicated manual data analysis
iQue Qbeads® Plexscreen are a family of reagents that allow researchers to analyze secreted proteins through their capture on distinct bead types, enabling multiplexed quantitation of biological parameters. Customers can choose from over 50 analytes to quantify human, mouse or rat secreted proteins or use iQue Qbead® Devscreen reagents to allow the flexibility to attach their own capture antibodies or target proteins. iQue Qbeads® are highly reproducible, significantly reduce sample requirements (as low as 2 µL) and time to results with less hands-on time and a no wash assay. The iQue Qbeads® assay can be multiplexed with other iQue® kits, for high-throughput measurement of protein secretion in addition to viability, marker expression and proliferation through a simplified, streamlined workflow. Save precious samples, time and get real time pharmacological analysis using integrated iQue Forecyt® software. Quantifying cytokine and signaling molecule secretion in adherent or suspension cells has never been easier!
Perform high-throughput analysis - Facilitate screening and profiling studies with rapid quantification of secreted proteins in 96 or 384-well plates.
Multiplex quantification of cytokines - Analyze up to 30 secreted protein concentrations in a single sample using customized or pre-configured panels.
Simultaneously quantify cytokines and cell readouts - Multiplex with other iQue® kits and/or antibodies for combined analysis of cell functions and subsets.
Maximize your productivity - Reduce reagent costs and time to answer – use as little as 2 µL in a simple, no wash protocol.
Perform high-throughput analysis
Enable screening and profiling studies with rapid quantification of secreted proteins in 96 or 384-well plates
Figure 1. Quantify cytokine secretion in minimal time using a 384-well assay format.
PBMCs (80K/well) from multiple donors were seeded alone (single donor controls) or in combination (8 donor pairs in total) in a Mixed Lymphocyte Reaction (MLR) assay. Cells were treated with several concentrations of an anti-CTLA4 checkpoint inhibitor antibody. Heatmap representation of secreted IFNγ across the 384-well assay plate. High speed analysis of a 384-well plate completed in 20 minutes.
Multiplex quantification of cytokines
Analysis of up to 30 secreted protein concentrations in a single sample using customized or pre-configured panels.
Figure 2. Analyze data quickly and easily using the iQue Forecyt® software system.
Dendritic cells were seeded with encoded CD4+ T cells (3:1 T cell-to-DC ratio) and treated with Pembrolizumab before being analysed on Days 2 and 6 using the iQue Qbeads® Human Inflammation Panel Kit. Pre-defined analysis templates are provided with each kit for the included analytes. (A) Gating template used to separate Qbeads based on their unique fluorescent signature. (B) Baseline secretion of multiple inflammatory cytokines measured using Qbeads in a single well performed in duplicate. (C) Efficiently present clear data trends using iQue Forecyt® heatmaps and concentration response curves of secreted proteins and cytokines.
Simultaneously quantify cytokines and cell readouts
Multiplex iQue Qbeads® with other iQue® kits and/or antibodies for combined analysis of cell functions and subsets.
Figure 3. Multiplexed quantification of secreted signaling molecules and cellular functions.
T-cells were activated using either Dynabeads or CD3-CD19 BiTE (Bi-specific T-cell Engager) and the production of cytokines was captured and analysed using iQue Qbeads® Plexscreen in addition to cellular viability (using the iQue® Cell Membrane Integrity Kit) within the same well. Lower levels of cytokine production in the BiTE treated t-cells compared to Dynabeads correlated with increased target cell killing. (A) Heatmap showing secreted concentrations of TNFα with increasing concentration of t-cell activators. Concentration response curves of IFNγ secretion after activation of t-cells with Dynabeads (B) and BiTE (C). Cellular viability with Dynabeads (D) and BiTE (E).
Maximize your productivity
Reduce reagent costs and time to answer – use as little as 2 µL conserving precious sample with no wash protocol.
Figure 4. iQue Qbeads® Plexscreen kit workflow.
(A) Schematic showing the simple workflow for measuring analyte concentrations using iQue Qbeads® Plexscreen. (B) The iQue Qbeads® assay builder provides a flexible assay format which facilitates analysis of up to 30 secreted protein concentrations in a single well. (C) Distinct identification of iQue Qbeads® and cells characterized by iQue Forecyt® software population gating.
Frequently Asked Questions
iQue QBeads® allow for the quantitative measurement of over 50 analytes, including both cytokines and chemokines. A single supernatant sample (10 µL) can be used to analyze up to 30 secreted proteins simultaneously which can be carried out for quantification of human, mouse and rat secreted proteins.
If your particular analyte is not offered in our current portfolio, iQue Qbead® Devscreen reagents allow the flexibility to attach your own capture antibodies or target proteins for a more bespoke, personalized solution.
Currently, iQue QBeads® have not been validated for use in the detection of intracellular cytokines.
iQue Qbeads® PlexScreen: Human Secreted Proteins
sCD154 (sCD40 Ligand)
CD178 (Fas Ligand)
CD121a (IL-1 RI)
CD121b (IL-1 RII)
iQue Qbeads® PlexScreen: Mouse Secreted Proteins
iQue Qbeads® Human Inflammation Panel Kit
|Platform: Compatible with iQue® 3/iQue® Screener Plus - BR and VBR Configuration|
1 x 384 wells
5 x 384 wells