01/23/2023 | Press release | Distributed by Public on 01/24/2023 11:43
ELISA has been the premiere immunoassay technique for decades. But it is problematic for the simultaneous detection of multiple analytes, especially at ultralow concentrations. This kind of detection is now necessary for the accurate and timely clinical assessment of complex diseases or biological processes, involving many biomarkers, from dilute, peripheral biofluids.
Automated assays that simultaneously detect and quantitate multiple analytes at ultralow concentrations are now commercially available. These "multiplex assays" are taking immunoassays to the next level of performance, beyond the limitations of conventional ELISA. Here, you'll see the current state of the multiplex assay in ELISA.
While conventional ELISA remains the workhorse for the detection of single analytes in small-scale settings, there is growing demand from research and the clinic for a reliable system for multi-analyte detection and quantification in complex biological processes. Neurological diseases, autoimmune diseases, and cancer require the identification and monitoring of signature cohorts of trace biomarkers. In this context, conventional ELISA can present a number of issues:
Efficient use of time, effort, and physical resources is crucial to modern research. Conventional ELISA's one analyte detection protocols consume valuable time, effort, reagents, and samples, while limiting throughput and accurate, timely analysis. Today, large cohorts of clinical or drug screening samples demand much more efficient, sensitive, accurate, and versatile immunoassay techniques to assess their inherent biological complexity.
Several varieties of multi-analyte detection array exist today. Planar arrays feature multiple capture and detection antibodies on a two-dimensional format. Flow cytometry-based analysis employs particles bearing reaction sites that can be read via LED or laser light as they file past a detector. Finally, multiplex bead array assays (MBAAs) are immunoassays that use capture beads as their reaction centers. In Simoa® assays , analyte-bound beads get easily parsed to individual wells and sensitively detected via fluorophores, emitting differential wavelengths associated to the beads. Other bead-based technologies maintain a mixed suspension of analyte-bound beads through detection, such is the case with cytometric-based detection methods.
Simoa® MBAAs offers many advantages over traditional ELISA:
In a further contest of multiplex assays vs ELISA, a recent publication1 compared with performance of high sensitivity ELISA against platforms from 3 commercial multiplex assay manufacturers, including Quanterix's Simoa® magnetic bead-based array assay.
Each system measured specific cytokine markers (IL-1 beta, IL-6, IFN-gamma, and TNF-alpha) in patients with Post-Traumatic Stress Disorder or Parkinson's disease. Healthy volunteers and clinical populations donated plasma and serum samples. The study characterized the detection limits and precision of each system through several key parameters:
Of all the platforms tested, Quanterix's Simoa® technology had the best sensitivity across all four cytokines for both the plasma and serum samples, exhibited the highest inter- and intra-assay precision, and had the highest sensitivity and precision across all four cytokines.
To quote the authors….
"Overall, the single molecule array (Simoa®) ultra-sensitive platform developed by Quanterix exhibited the best performance across all cytokines and should be recommended for use when highly sensitive and precise immunoassays are required."
-Heather C.Lasseter, Allison C.Provost, Lauren E.Chaby, Nikolaos P. Daskalakis, Magali Haas, and Andreas Jeromin
Other examples and resources
Ready to take your immunoassays to the next level of performance?