Why do you study cytokine release after 4 hours and not 24 hours?

The blood in our platform remains in constant circulation, and your drug is added within minutes upon acquisition from the blood donor. We study cytokine release after 4 hours because this mimics the time point when a rapid infusion reaction (such as a cytokine release storm) occurs in the clinical situation – within a few hours of drug infusion. (NOTE: Other methods such as PBMC or WB plate-based assays may use the “4-hour” timepoint in their marketing, however, that is misleading as it commonly refers to how long their blood is stored in a buffy coat. In the marketing of such assays, it’s not uncommon to leave out that the incubation time is at least 24 hours in order to detect any response at all – this does not mimic the clinical situation)

Can you analyze cells after they are run in your assay?

Yes. We have great experience in staining and analyzing blood cells (RBCs, PLTs, granulocytes, DCs, monocytes, NK cells, B cells and T cells) using flow cytometry.

Do the regulatory authorities accept Immuneed’s method as a cytokine release assessment?

Yes, they do.  Our clients have included the results from Immuneed in their IND/CTA filings to regulatory authorities, without positive results.

What guidelines do the regulatory authorities have for biological drugs regarding cytokine release?

In order to mediate cytokine release and cellular activation, regulatory agencies recommend in vitro assessments of such products in human whole blood or peripheral blood mononuclear cells, so as to overcome the known limitations of animal models. (Ref: FDA, Guidance for industry, Immunogenicity Assessment for Therapeutic Protein Products, August 2014.)

What drugs can you study?

We can study biological drugs, advanced-therapy medicinal products and small molecules, as well as nanoparticles.

What kind of blood do you use?

We can utilize both patient blood and healthy blood. If a client desires, we recruit healthy volunteers and analyze blood-drug interactions at a preclinical stage. These healthy volunteers are the same people that are enrolled in the subsequent clinical trial.

What patient groups can you recruit?

We can recruit patients with B-cell malignancies or multiple sclerosis.

What are the main benefits of Immuneed’s platform compared to PBMC assays or whole-blood plate assays?

We collect blood with a method that keeps all the cells intact and viable. Since the blood is kept in circulation, and the analyze is initiated immediately (not stored), we do not need to add high concentrations of anticoagulants that affect your test result. PBMC assays and plates with whole blood often fail in predicting responses to substances known to cause severe cytokine release in humans. Not only can Immuneed’s platform detect that a substance has a high risk of inducing cytokine release, but it can also capture moderate to low responses. We have conducted a comparative study to demonstrate the benefits of our platform. You can find the study here.

Which cytokines can you measure?

We can measure the standard proinflammatory cytokine panel including cytokines of the adaptive and innate arms: IFNg, IL-2, IL-6, IL-8 and TNFa.

Is the complement system still active even though Immuneed use heparinized tubes for the circulation of blood?

Yes, it is.

How do you prevent clotting?

Our platform ensures that the blood is kept in constant circulation, similar to the way it would be in-vivo. This keeps clotting from occurring.

How many blood-loops can you run in parallel?

We can run 25–30 loops in parallel per donor.