Cytokine Release

Assessment in circulating human whole blood.

Leverage the benefits of the unique test system ID.Flow®


We draw blood from donors directly at our test site allowing us to observe the effects of drugs on fresh, minimally manipulated whole blood, offering you invaluable insights into how drugs affect complex biological systems in the human blood.


ID.Flow is a highly advanced test system that ensures the continuous circulation of human blood and test substance at a regulated temperature of 37°C. Its superior sensitivity and dynamic features set it apart from PBMC- and whole blood assays.


FDA and EMA encourage the use of New Approach Methodologies (NAMs) to evaluate the immunotoxicity potential of new drugs. ID.Flow is a NAM that is accepted as a cytokine release method by regulatory authorities.

Cytokine Assay

Unmatched sensitivity in grading cytokine release.

Regulatory authorities require an assessment of cytokine release potential, emphasizing the importance of using human-based methods to determine potential immunotoxicity: Immuneed’s innovative test system, ID.Flow  offers unmatched sensitivity in grading cytokine release. By assessing cytokine release in fresh human whole blood directly upon blood draw, ID.Flow detects immune responses that can be difficult to determine in in vitro plate-based methods and animal models.

ID.Flow exhibits minimal background and significantly less donor variability.

The figures above shows a comparison between ID.Flow and two plate-based assays that used either PBMCs with coated antibodies or whole blood with soluble antibodies to assess cytokine release. The antibodies used in the tests were classified as having a low risk of causing CRS, such as anti-integrin alpha 4 (1 µg/ml) and anti-EGFR antibody (200 µg/ml), and high risk of causing CRS, such as anti-CD52 antibody (3 µg/ml) and anti-CD28 (aCD28). The level of cytokines was compared to the vehicle and zero (blood directly after acquisition). Readouts from ID.Flow were taken after 4 hours which mirrors the actual time for cytokine release upon IV-administration in humans, whereas the plate-based assays required a 24-hour incubation period to distinguish a positive response from a negative.

The data from the comparative study demonstrate that ID.Flow has a superior ability to detect cytokine release. This is shown by the greater signal-to-noise ratio, i.e. greater separation of cytokine levels induced by the high risk antibodies than the low risk antibodies , which makes it possible to detect not only super-agonistic responses but also low-to-moderate responses. 

ID.Flow cytokine assessment

  • The freshly donated human blood is immediately transferred to the ID.Flow test system, followed by administration of the test items, and set to circulate at 37°C to prevent clotting.
  • Samples are collected at baseline and selected timepoints, then processed to plasma by centrifugation.
  • Cytokine analysis is conducted on the Multi-Array platform from Meso Scale Discovery (MSD).


  • 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.
  • All available analytes for the MSD platform can be included in the study.

Read more about how ID.Flow was used to test a TGN1412-like antibody, Fletcher Int Immunopharmacol. 2017 Oct 27;54:1-11. follow the link.  (ID.Flow = whole blood loop system in this publication)

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Product comparison: cytokine release assays

We evaluated the predictive value of different cytokine release methods. Download our white paper to learn about thr pros and cons of three different methods of assessing cytokine release.