Science-led approach - optimization
Our tailored and customized approach ensures a robust experimental design to provide a strong data package and address your scientific questions.
- Two methods of Test Substance exposure to the biological sample (PBMCs and Whole Blood) are assessed – wet coated onto a plate or applied in aqueous phase.
- 44 groups for optimization including reference controls Lemtrada® (Alemtuzumab) and Lipopolysaccharide (LPS) known to cause cytokine storm in patients.
- Th1/Th2 cytokine release profiles are measured using the Multiplex Luminex platform. An exemplar cytokine panel includes GM-CSF, IFN-γ, IL-β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNFα.
Methods of data analysis
For each sample replicate the fold change over vehicle control is calculated, a 2-fold change over vehicle control is considered as relevant. The percentage of responding donors can also be calculated and compared to relevant reference controls.
Using standard assay reference control, Lemtrada® and LPS induce a consistent and widespread cytokine release effect across the donor cohort.
How our cytokine release assay is performed
|1. Peripheral blood samples from multiple donors are used to ensure genetic diversity.||2. Test and control compounds are prepared to required concentrations and applied wet coated onto assay plates or used in aqueous phase; diluted blood is then added and plates are incubated for 1, 6 or 24 hrs.|
|3. Test substances interact with their specific cellular targets leading to the release of cytokines from activated immune cells.||4. Assay plates are spun down and supernatants are used to measure a panel of cytokines such as GMCSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNFα using the MSD technology.|
|5. Data are analyzed and displayed in a series of convenient formats such as heatmaps, bar graphs and dot plots to provide a quick overview of cytokine responses across different donors or to determine the population of responders vs non-responders for individual treatments and cytokine.|