Cytokine Detection and Measurement with Multiplex Technologies

Posted on by Luminex Corporation

Cytokines are molecules that immune system cells use to send signals to each other. They are of particular importance for the recruitment of immune system molecules to the site of an invading pathogen.

The discovery and identification of cytokines is a crucial part of biological research – they provide insight into physiological processes and disease pathways, and can serve as biomarkers for various diseases. They are classified as proteins, glycoproteins, and peptides depending upon their individual characteristics.

Multiplexing technology has become an important tool in cytokine detection. Assays can be performed with great speed and accuracy by making use of hundreds of specially prepared magnetic beads, or microspheres. The microspheres are dyed internally with a mixture of dyes to provide a unique spectral address – when mixed with a sample, molecules on the outside of the spheres react to any molecules that they are designed to capture. A detection molecule is then added and they are run through an instrument that determines the spectral address of the microsphere and quantifies the signal of the captured molecules, thus reporting on the contents of the sample.

The value of multiplex analyses has been confirmed by research from the Southwest Foundation for Biomedical Research into cytokine detection. This work, completed by Luis D. Giavedoni, concluded with the creation of an assay that can be used to detect 20 non-human primate (NHP) cytokines and chemokines.

These cytokines and chemokines include G-CSF, GM-CSF, IFN-gamma, IL-1beta, IL-1Ra, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p40), IL-17, IL-18, MCP-1, MIP-1alpha, MIP-1beta, RANTES, TNF-alpha, and TNF-beta.

Although this research was done with NHPs, the results can still have important repercussions for human health. Many physiological mechanisms are the same or similar between primates – work like this helps researchers understand just how similar they are and how models of disease in NHP can be translated to humans. The newly created assays can be used to develop better models of animal disease and vaccine development.

Another study, conducted at the University of California at Irvine, compared the use of the traditional cytokine assay, known as ELISA (enzyme-linked immunosorbent assays) with the more recent Luminex xMAP®, a multiplex assay method. The researchers studied samples from 96 pregnant women for cytokine concentration using each method.

They found that in the majority of cases the Luminex xMAP Technology is very useful compared to ELISA. Luminex Technology:

  • Makes use of smaller sample volumes
  • Delivers a result that is just as accurate
  • Processes with a higher throughput
  • Costs less

The study concludes, “Luminex multiplex technology has distinct advantages and is a valid alternative method to ELISA for the evaluation of the majority of cytokines tested and for the characterization of immune system status.”

The massive complexity of physiological molecules and processes require highly accurate, highly efficient methods of assay. Luminex multiplex technologies provide a vastly enhanced method of cytokine discovery and identification, allowing for the testing of dozens of cytokines at once, from a single sample. High-quality multiplex technologies have been proven to be at least as accurate as traditional methods while being much more time- and cost-efficient, and they are accelerating medical research in numerous ways.

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