Multiplexed ELISA: The Future of Lab Testing Revealed!

Over the last 20 years, biological assay development has evolved through the advent of high throughput technology. Such technological advancements have helped us decipher numerous pathophysiological processes, paving the way to notable impacts on biotherapeutics and diagnostic applications. Multiplex ELISA is one such robust platform for accelerating drug development. 

Although singleplex ELISA is sufficient to accurately diagnose diseases involving the characterization of a single analyte, monitoring complex and multifactorial diseases requires multiplex capacities. Accordingly, ELISA development for incorporating novel biomarkers to study multiple analytes in a single patient reduces assay time, cost, and sample volume needed for bioanalysis. Moreover, multiplex ELISA assays are helping researchers move towards personalized and precision medicine. Therefore the current article focuses on multiplexed ELISA. However, a thorough ELISA validation will remain crucial for generating accurate, reliable, and reproducible study results. 

Why is Multiplexed ELISA the Future of Lab Testing?

Over the past 50 years, immunoassays have been  the gold standard  for drug development studies. Advances in hybridoma technology for generating monoclonal antibodies facilitate the development of robust and accurate antibody-based immunoassays. Although singleplex and multiplex ELISA both employ sandwich format, singleplex ELISAs are usually based upon colorimetric reaction. On the other hand, multiplex ELISA uses fluorescent and electrochemiluminescent reporter systems, which are more compatible with simultaneous analysis.

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Multiplex ELISA assays can be divided into two formats: planar and microbead-based suspension assays. The planar format includes assay platforms such as qPLEX, array, and MSD multiplex ELISA assays. And the other hand, the bead suspension assay format includes BioPlexPro, cytometric bead arrays, and the Luminex platform. However,  multiplexed ELISA requires extensive qualification and validation processes compared to the singleplex format. Let us explore some trending multiplex immunoassays

The applications of multiplex ELISA assays have resulted in the development of numerous in vitro diagnostics and point-of-care testing tools. Over the last decades, several multiplex ELISA formats have demonstrated value in research and clinical studies. Multiplex assay formats can substantially  improve disease diagnosis, monitoring, and management. However, most multiplex assays require expensive setups for bench-top readers. Cost-effective point-of-care tools can prove beneficial in real-world uses. Besides, emerging technology in the smart digital healthcare space facilitates advances in personalized healthcare. 

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A prospective multiplex format uses a solid substrate-based spot microarray to detect multiple analytes in a single study sample simultaneously. Spot microarrays often require expensive and complex preparation. Besides, they adopt a traditional manual ELISA protocol. 

The xMAP technology is one of the widely used multiplex formats. This technology can detect up to 100 target analytes in a single well. It is one of the most robust multiplex immunoassays available for bioanalysis. However, it requires expensive instrumentation. On the other hand, the ELISA format developed by Meso Scale Diagnostics is another exciting multiplex format to detect analytes in complex biological samples. This format uses carbon electrodes in microplate surfaces to capture the analyte of interest. The primary advantage of MSD multiplex ELISA assay is its analytical performance and less labor-intensive experimental protocol. 

In conclusion, multiplexed ELISA has the potential to accelerate drug development phases and is proving critical for the future of lab testing. 


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