Multiplex cytokine assays can help understand the underlying mechanisms of several biological and pathological processes. Luminex multiplex bead-based assays are immunoassays that measure multiple analytes in a single assay volume. This technology employs Luminex xMAP beads to detect up to 100 analytes in a single sample simultaneously. Color-coded beads are dyed with different proportions of fluorophores, where each fluorophore corresponds to a distinct spectral signal.
Luminex analysis has several advantages.
- Conserves sample by assessing multiple analytes in less than 25 μl of sample
- Saves time and resources
- Allows the flexibility of building your panel
- Reduces experimental variability
The current article is a brief on Luminex cytokine assay. It discusses the profiling of multiplex immunoassays for cytokine analysis.
Profiling of bead-based assay
Bead-based assays begin with coupling analyte-specific antibodies to unique bead regions. Once these antibodies are captured on the plate, they are incubated with the sample. After the washing step, biotinylated detection antibodies and the reported molecule are introduced into the incubated mixture. The detection instrument has two sets of lasers. The first laser excites the beads to identify the analyte-specific bead region, while the other laser determines the generated signal.
Bead-based assay software offers a variety of standard curves that may fit data analysis. The mean fluorescent signals generated by the bead-based assays are directly proportional to the amount of analyte captured by the beads. Corresponding levels of analytes are measured using standard dilution curves. Such features help identify the best-fit signal for the standards and sample concentrations.
Multiplex immunoassays are ideal for assessing biomarkers in complex matrices. They may be even more significant in complex diseases such as HIV. HIV patients are susceptible to infections. Studies have identified numerous biomarkers associated with HIV infection. Identifying biomarkers in these patients can help clinicians monitor and segregate patients in different stages, such as incubation, acute, convalescent, and recovery phases.
Moreover, multiplex immunoassays simplify the determination of analyte profiles. Multiplex assays require a single sample to analyze multiple targets. This feature is particularly advantageous with limited sample volumes such as CSF and pediatric samples. Bead-based assays offer good precision, higher sensitivities, and a broader detection range. Furthermore, smaller sizes and virtual liquid analysis offer faster assay kinetics.
Due to their multiplexing capacities complemented with accuracy, speed, and specificity, multiplex immunoassays have become the primary choice of bioanalytical technique in protein analysis. As this technology is adaptable to cytokines, chemokines, and other immunomodulatory elements, multiplex cytokine assay are widely used to evaluate the immune response and immune pathogenesis.
Numerous studies have investigated cytokines using multiplex assays. These assays can monitor immune responses against several infectious diseases. For instance, sepsis and infection are major causes of neonatal mortality and morbidity. Early diagnosis of these diseases is crucial. However, there are several challenges as the clinical signs are subtle and highly variable. Advances in pediatric infection and diagnosis have been slow, but advances in multiplex analysis have shown promising results in early diagnosis of sepsis and infection.
In conclusion
multiplex cytokine assays have skyrocketed the science of cytokine analysis.
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