Cytokines are cell signaling molecules modulated by the immune system. They are mainly associated with cell growth, differentiation, and proliferation. Moreover, cytokines regulate various biological functions, including inflammation, innate and adaptive immunity, angiogenesis, tumorigenesis, immune cell differentiation, neurobiology, and pathogenesis. Studies have shown that dysregulation in cytokine expression is related to several inflammatory and immunological diseases. Usually, immune cells release cytokines to attract cells of the immune system to the site of injury or infection.
ELISA assays are the most common bioanalytical method validation employed to study cytokine expression. However, it is crucial to understand that instead of evaluating a single cytokine, assessing a cluster of cytokines gives a better picture of the disease mechanism. Hence, multiplex cytokine assays are a better option in studying cytokine expression. Several multiplexing platforms are available for evaluating cytokine expressions. These platforms include Luminex-based assays and Meso Scale Discovery (MSD) assays. Both these formats have their advantages. So let us explore the role MSD and Luminex play in unraveling complex diseases.
Multiplex cytokine assays for complex diseases
Cytokines are cell signaling molecules, and as mentioned earlier, they are crucial in recruiting immune cells in case of infections and injury. Cytokine discovery and identification have become critical components of biological and pharmaceutical research. Cytokines can either be proteins, peptides, or glycoproteins, depending on their environment and their individual properties.
Multiplex cytokine assays are robust methods for generating extensive quantitative data from a single assay sample. The assay volume required per sample is less than 50 microlitres and hence is advantageous with limited sample volumes. As MSD and Luminex cytokine assays can measure multiple analytes in a single sample, they expand our current understanding of disease states and mechanisms of drug action.
Multiplexing capabilities mean assays can be performed at high speed with reliable delivery of results. Let's take Luminex multiplex assays as an example of multiplex technology. Luminex assays use specially designed magnetic beads, called microspheres. These magnetic beads are internally dyed with different proportions of red and infrared dyes to generate unique spectral signatures. These beads react with the analyte of interest and capture them accordingly. A detection molecule then detects the conjugates, and depending on the spectral signature, quantifies the analyte of interest. Moreover, custom Luminex assays provide the opportunity of choosing and designing the assays to match several analytes of interest.
Today more studies are employing multiplexing strategies for routine cytokine assays. Not only with non-human samples, but multiplexing with human samples is increasingly used in clinical and non-clinical studies. The basis of understanding a cytokine expression is in the ability to assess multiple cytokines together. Hence, multiplex cytokine assays have become the obvious choice in understanding complex disease mechanisms.
The road ahead
Today it has become evident that to comprehend the complexity underlying several disease processes, researchers should discover highly efficient and reliable assay methods. Multiplex cytokine assays provide enhanced technological solutions for identifying and quantifying multiple cytokines in a single study sample. In summary, multiplex cytokine assays have played a substantial part in accelerating clinical and non-clinical research.