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384-Well Multiplexed Luminex Cytokine Assays for Lead Optimization
Huaping Tang1, Reshma Panemangalore1, Melissa Yarde1
1Bristol-Myers Squibb, Lawrenceville, NJ, USA.
Insights
A new miniaturized Luminex assay enables high-throughput, multiplexed measurement of cytokines in whole blood. This cost-effective platform accelerates drug discovery and biomarker profiling for various diseases.
Area of Science:
- Immunology
- Biotechnology
- Assay Development
Background:
- Cytokines are crucial signaling molecules in immune responses and disease pathogenesis.
- Abnormal cytokine levels are implicated in chronic inflammation, autoimmune diseases, and cancer.
- Current cytokine quantification methods like ELISA are costly and low-throughput.
Purpose of the Study:
- To develop a miniaturized, high-throughput Luminex assay for multiplexed cytokine measurement.
- To reduce reagent consumption and costs compared to traditional methods.
- To enable efficient biomarker profiling in translational research.
Main Methods:
- Development of a miniaturized 384-well Luminex assay platform.
- Utilized <25% of reagents compared to standard 96-well assays.
- Validated the assay for measuring cytokines in human whole blood.
Main Results:
- Established a high-throughput, multiplexed cytokine assay panel.
- Demonstrated reduced reagent usage and cost-effectiveness.
- Enabled high-throughput screening of compounds and biomarker profiling.
Conclusions:
- The miniaturized Luminex assay offers a scalable and efficient solution for cytokine analysis.
- This technology facilitates high-throughput drug discovery and biomarker profiling in translational research.
- The platform supports analysis of biofluids from preclinical and clinical studies.
Abstract:
Cytokines serve as a major mechanism of communication between immune cells and are the functional molecules at the end of immune pathways. Abnormalities in cytokines are involved in a wide variety of diseases, including chronic inflammation, autoimmune diseases, and cancer. Cytokines are not only direct targets of therapeutics but also important biomarkers for assessing drug efficacy and safety. Traditionally, enzyme-linked immunosorbent assays (ELISA) were most popular for identifying and quantifying cytokines. However, ELISA is expensive, labor intensive, and low throughput. Here, we report the development of a miniaturized Luminex (Austin, TX) assay platform to establish a panel of high-throughput, multiplexed assays for measuring cytokines in human whole blood. The miniaturized 384-well Luminex assay uses <25% of the assay reagents compared with the 96-well assay. The development and validation of the 384-well Luminex cytokine assays enabled high-throughput screening of compounds in primary cells using cytokines as physiologically relevant readouts. Furthermore, this miniaturized multiplexed technology platform allows for high-throughput biomarker profiling of biofluids from animal studies and patient samples for translational research.
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