Comparative Analysis of Different Cytokine Assay Methods

Uncover the best cytokine assay method for your research. Compare MSD and GLP Lab techniques in detail. Learn about their advantages, limitations, and applications.

Comparative Analysis of Different Cytokine Assay Methods

Cytokines are small secretory proteins that play a crucial role in cell signaling. Cytokines include interferon, interleukin, growth factors, tumor necrosis factors, and chemokines. Cytokines bind to the corresponding receptors on an effector cell and begin signal transduction. It is well-known that cytokines are critical in several immunological responses, such as infection, tissue repair, inflammation, homeostasis, hematopoiesis, and cellular growth and development. Dysregulation or dysfunction of cytokines is associated with multiple diseases. Hence, it is crucial to study cytokine regulation to understand disease mechanisms and identify novel drug targets. 

As cytokines have crucial biological functions and considering their role in diseases, cytokine profiling techniques are necessary for drug discovery and development. Sensitive and robust cytokine assay technology is required for GLP labs to facilitate reliable drug candidate screening and disease biomarker identification. The current article explores different cytokine assay methods. 

Cytokine bioassay

Bioassays evaluate cytokines within a system by assessing their biological activity and associated readout. In bioassay, the sample activity is evaluated using a sensitive cell line, and the data is compared to standard cytokine results. This type of cytokine assay is sensitive and allows the identification of bioactive molecules in a sample. 

Cytokine protein microarrays

Protein microarrays evaluate the functional activity and interaction of cytokines on a larger scale. This method is sensitive and offers high throughput assessment, allowing the estimation of multiple cytokines in parallel. This assay uses a protein platform with a surface containing different capture proteins. Labeled molecules are then introduced to this array, and after interaction with the capture protein, a signal is generated and read by the scanner. Analytical microarrays use aptamer, antibodies, or affibodies at the chip surface to bind the cytokine of interest within a protein solution. 

High-performance liquid chromatography

High-performance liquid chromatography identifies cytokines based on their specific weights, ligand affinity, ion exchange, and hydrophobicity. Hence, HPLC assays exploit specific chemistries of the target compound to separate and identify cytokines in a study sample. High-performance liquid chromatography has successfully purified and separated cytokines such as IL-1 from different cells, for example, macrophage and epidermal cells.

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Sandwich antibody assays

For an assay to be ideal for cytokine analysis, it should meet several performance parameters. However, assay formats such as protein microarrays and HPLC systems offer suboptimal performance or conflicting results during cytokine analysis. Two sandwich antibody assay formats, a plate-based and a bead-based assay, are ideal assay formats for cytokine analysis. 

Plate-based assay formats such as ELISA and MSD cytokine analysis and bead-based formats, for example, multiplex immunoassays, are widely used for cytokine biomarker discovery. These assay formats follow the principle of sandwich assays, where the cytokine of interest is sandwiched between two specific antibodies that bind to non-competing cytokine epitopes. In bead-based assay format, the antibodies act as a capture antibody by coating to a solid bead or act as a secondary antibody where it is bound to a reporter molecule. On the other hand, in a plate-based sandwich assay, capture antibodies are fixed at distinct points within an assay plate. 

In conclusion

Cytokines are vital cell signaling molecules. Hence, researchers must have a complete understanding of different cytokine assay methods. 

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