Cytokine Release Assay

Assessment in circulating human whole blood

Customer promise


Ex-vivo assessment that helps you understand your drug’s potential to mediate cytokine release and cell activation. Our cytokine assay offers unmatched sensitivity in evaluating the potential for adverse pro-inflammatory reactions.


ID.Flow provides a unique and physiologically relevant testing environment. Using circulating whole blood, we detect responses frequently missed out by plate-based assays.


Our Scientific Advisors will design the study to suit your unique project. Choose between a scientific reporting format that suits your project: Standard Report IND-enabling Report

Cytokine Assay

Cytokine release – a critical aspect of preclinical safety assessment.

Regulatory authorities demand an assessment of cytokine release potential, emphasizing the importance of human-based methods in determining potential immunotoxicity. Leverage the benefits of CRA in Immuneed’s innovative platform, ID.Flow. It offers a sensitive prediction of cytokine release in an environment that closely mimics human blood circulation.

Traditional plate-based assays require high levels of anticoagulants which increase the risk of a missed response by inactivating complement and cascade systems. In contrast, ID.Flow utilizes fresh whole blood with all cells and systems present and active. ID.Flow can provide readouts within minutes to hours. This perfectly mirrors the clinical situation, where reactions develop swiftly after drug administration and not following a lengthy incubation period. With the high sensitivity and speed of our method, you can effectively assess the hazards of cell activation and cytokine release posed by your drug candidate.

Schedule a meeting with us to discuss your project and we can tailor the study to address your specific needs.

The analytic power of ID.Flow

Shown above is a direct comparison of Immuneed’s platform, ID.Flow with two plate-based assays where either PBMCs with coated antibodies or whole blood with soluble antibodies were used for cytokine release assessment. It involves antibodies associated with a low risk of causing CRS, namely anti-integrin alpha 4 (1 µg/ml) and anti-EGFR antibody (200 µg/ml), as well antibodies associated with a high risk of CRS, namely anti-CD52 antibody (3 µg/ml) and anti-CD28 (aCD28). The level of cytokines as compared to vehicle and zero (blood directly after the acquisition). In the ID.Flow, cytokine release was measured at a time point of 4 hours, while the plate-based assays required a 24-hour incubation time before readouts could be taken.

Read more about when Immuneed’s platform was used to test a TGN1412-like antibody, Fletcher Int Immunopharmacol. 2017 Oct 27;54:1-11. follow the link.  ID.Flow is called whole blood loop system in this publication.

Analysis by ID.Flow

Your drug is infused into fresh human blood in circulation.
Read-outs are taken to perfectly mimic the clinical situation of infusion reactions.
The multiplexing platform from MesoScale Discovery (MSD) is used for analyzing the samples.
All available analytes for the MSD platform can be included in the cytokine evaluation if you wish.


The standard proinflammatory cytokine panel includes cytokines of the adaptive and the innate arm: IFN-γ, IL-2, IL-6, IL-8 and TNF-α.  Other readouts are available upon request.

Talk to us!

Have a project or a question, or would you like to request a quote? We'd be happy to hear from you.

White Paper

We have evaluated the predictive value of different cytokine release methods. Read our white paper to discover the pros and cons of the different assays.