I have plasma samples from patients and controls which I would like to assay for cytokines including IFN-g, TNF, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-21as well as granzyme A and B. Is there a multiplex assay which is sensitive enough to do this?
Hi there! I had very good experiences with multiplex assays from Mesoscale Discoveries (MSD). They offer a range of electrochemiluminescent cytokine multiplex assays which in part can be configured by the customer. Essentially, bead-based arrays, like Luminex, may be used as well. For both, you will need special equipment, which is affordable, if you do not chose the high-end readers. Sensitivity of individual markers should be discussed with the suppliers, but your choice of markers does not seem to include very exotic ones with extremely low abundancy.
(I am not affiliated to MSD, or have any advantages when recommending their products! I only was very satisfied with the product range and the service)
Hi Emily - what are your cohort sizes and how many cohorts do you have?
best
G
Hi Emily,
The Bioplex bead assay is a very handy tool as it uses little of your sample and can give you up to 28 cytokine levels. Not cheap though and takes a bit of tweaking. Experience shows that many of the patient samples will have low/non-detectable levels ...
regards, Hans
Thanks everyone.
Grant - At the current moment I have 2 cohorts of 12 that I plan to analyse.
Hi Emily,
I have to agree with Just Genius. I have run tens of thousands of plasma samples on the MSD platfom. We have run many comparisons, you just can't beat them for consistency of results.
In my experience the only way to reliably test the bioavailable levels of IL-2 is by the in vivo Cincinnati capture assay. Working with toxoplasma peak infection it was never detectable. Other cytokines should be fine.
Hi, Emily, Mulitplex kits from Biorad and Millipore works nicely for most of those cytokines.
Thanks everyone. Does anyone have any idea what levels I am looking at for any of these cytokines in healthy patients?
I had good experiences with multiplex kits (magnetic beads) from Millipore.
Cat. No. HCYTMAG-60k-PX30 (IFN-gamma, TNF alpha, IL2, IL4,IL5, IL6,IL9, IL10, IL13, IL17A and others). Also look the kits HCP2MAG-62k-PX23 (for IL-21) and HCD8MAG-15k (for Granzyme A and B).
I use MSD and I am very pleased with them. Very sensitive (3 logs), low volume (25ul). Can be customized in term of what cytokines you need to measure. However, you will need to purchase their reader. They have several models but they are pricey. They do have special prices for academia. You can certainly find somebody with such reader who will be willing to collaborate with you.