Simultaneous measurement of multiple cytokines
the simultaneous measurement of 12 human cytokines from human serum
samples enabling cytokine studies to be conducted faster.
Cytokines are small regulatory proteins produced by various cell types and function as intracellular chemical messengers and control many immune and inflammatory responses.
They are normally released in picomolar amounts, making their detection somewhat difficult and are involved in a variety of immunological, inflammatory and infectious diseases.
The new research, published in the Journal of Proteome Research , describes how a biochip array can be used to simultaneously measure 12 different cytokines from up to 54 different biological samples.
As such this new tool may enable researchers conducting clinical studies on diseases such as cancer, cardiovascular conditions, autoimmunity, nervous system dysfunction, ageing and sepsis to study the effects of drugs on these potential biomarkers of disease progression.
Indeed, the Northwick Park tragedy that occurred while researchers were conducting 'first-in-man' clinical trials on TeGenero's TGN1412, which instead of subtly 're-tuning' the immune system as the developer hoped sent the immune system into overdrive and caused a 'cytokine storm' that attacked healthy organs with tragic results.
While the new system would have not been able to help minimise the tragedy, it may have enabled researchers to gather data quicker and followed the progression of the problem more closely than the currently used ELISA (enzyme linked immunosorbent assay) tests.
The biochip uses a sandwich immunoassay in which monoclonal capture antibodies are bound and stabilised to predefined positions on a pre-activated biochip surface.
These antibodies specifically bind to either one of 7 interleukins (IL-1a, IL-1ß, IL-2, IL-4, IL-6, IL-8 or IL-10) vascular endothelial growth factor (VGEF), interferon gamma (IFN?), epidermal growth factor (EGF), monocyte chemotactic protein 1 (MCP-1) or tumour necrosis factor alpha (TNFa).
After the addition of a small amount of a human serum sample was added to the biochip, a series of incubation steps were required before the results could be read using Randox Laboratories' Evidence Investigator system.
The new assay acheived sensitivity ranging from 0.12pg per ml for the IL-6 analyte, to 2.12 pg per ml for IL-4.
"This approach reduces the sample / reagent volume consumed with the corresponding effects in the cost effectiveness and can provide new insights in the clinical relevance of cytokines as it enables simultaneous measurement of very low levels of these compounds in clinical samples," write the authors.