Cytokine Release Assay (CRA) Services

How will my Cytokine Release Assay readouts look?

We will present the data in an easy-to-understand and digestible format.

In the example (Figure 1), PBMCs were stimulated with various concentrations of ‘Antibody X’ or OKT3 (positive control), and the serum was subsequently harvested and assayed for IL-2 (1A), IL-4 (1B), IL-6 (1C), IL-10 (1D), TNF-α (1E), IFN-γ (1F), and IL-17A (1G). Here, symbols within each stimulation condition represent one individual donor, while the black line denotes the mean concentration of all 4 donors. In addition, the data is customized to your set of cytokines with each donor shown so you can observe the level of consistency from donor to donor. Of course, the data can also be presented in different formats depending on your requirements. Finally, we will help you interpret the data so you and your team can feel confident in making decisions regarding the next steps in the development of your molecule(s).

Figure 1: Mean serum concentration of IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ, and IL-17A from ‘Antibody X’ treated PBMCs.Through the entire process, you can be assured that iQ will provide the best quality in reagents, technical skills, and service for this critical preclinical assay. Find out today how iQ Biosciences can help achieve your goals the smarter way.

Cytokine Release Assay (CRA) Background

With the current trend of developing large or small molecule-based immune-modulatory therapeutics for various oncology and autoimmune indications comes the risk of unwanted immune side effects in first-in-human trials. At iQ Biosciences, we aim to provide drug developers with additional information about their molecule before proceeding to clinical trials. iQ provides immunogenic assays (like this Cytokine Release Assay) to screen for markers that may be predictive of dangerous side effects after infusion into humans.

One of the most dangerous side effects from the administration of a therapeutic molecule is a cytokine storm or cytokine release syndrome (CRS)1. After administration of the therapeutic, the patients experience a release of pro-inflammatory cytokines, which results in symptoms that range from fevers and rashes to more severe and life-threatening ones, such as cardiac arrest or organ failure. These are unwanted side effects of the therapeutic and can be predicted with Cytokine Release Assays we conduct as a service at iQ.

In the competitive field of therapeutic development, ensuring the molecule does not elicit a cytokine storm is critical. In addition, with the growth of companies specializing in biosimilars, as well as new technologies, such as alternative antibody formats, obtaining cytokine data via multiplexed cytokine quantitation is a necessary step toward a preclinical data package before in-human trials.
Accordingly, financial costs and, more importantly, risk to the patient’s health can have serious implications for the success of a therapeutic program. With iQ Biosciences’ expertise in cytokine profiling to assess the levels of a variety of cytokines, drug developers can use our predictive human cytokine data to help assess the direction of their potential therapeutics and research programs.

Importance of CRA testing (Cytokine Storms)

The most well-documented case of a biologic that caused a cytokine storm is TGN1412, an anti-CD28 “superagonist” antibody that was proposed to treat certain autoimmune diseases and hematologic cancers2.  The adverse events experienced by patients after administration included pain, amnesia, diarrhea, vomiting, fever, and severe multi-organ failure1.  More importantly, the cause of these life-threatening effects was later attributed to a cytokine release storm1.  However, the preclinical safety testing showed no evidence TGN1412 would cause such severe and adverse reactions in first-in-human trials.  While TGN1412 is an extreme case, approved anti-HER2 and anti-CD52 antibodies represent other therapeutic examples that have been associated with pro-inflammatory clinical infusion reactions2 (for more information, see our blog on Cytokine Release Assays).

Current Regulatory Agency Recommendations Regarding Cytokine Storms

The FDA strongly recommends performing experiments to predict cytokine storms in the preclinical testing phase before first-in-human studies. At iQ Biosciences, we have developed expertise to better predict cytokine storms with Meso Scale Discovery (MSD) technology and optimized cytometric bead arrays (CBA) that assess the release of cytokines from human PBMCs in response to therapeutic molecules, both in wet-coated and soluble treatment formats. Cytokine detection assays and methods are in concordance with European Medicines Agency (EMA) and FDA recommended guidelines, including measuring serum IL-2, IL-6, IL-10, IFN-γ, and TNF-α.