A multiplex test is a form of immunoassay used in the biological sciences that use magnetic beads to assess numerous analytes concurrently in a single experiment. An ELISA variant known as a multiplex assay uses beads to bind the capture antibody. The market for multiplex assays was valued at $3.36 billion in 2021 and is anticipated to grow to $6.47 billion by 2030, registering a CAGR of 7.53 percent from 2021 to 2030.
Several common and reliable tests for single analytes
For many years, the cornerstone of biological research has been assays that quantify the concentration of a single protein in a homogeneous liquid sample (usually ELISA or western blot). Whether measuring the quantities of a specific protein or checking for the presence of protein indicators, accurately quantifying protein levels is useful in research and diagnostics.
There are now several ELISA kits on the market that use optimised antibody pairs. These kits include capture antibodies that draw an interest target from the biological environment and a particular detector antibody. ELISA uses specific, sensitive antibody pairings that can be read using common plate readers.
The Multiplex immunoassays
Methods that integrate tests for several target analytes are needed to solve workflow and sample volume issues. Multiplex immunoassays reduce workflow and sample volume issues by combining tests for numerous target analytes in a single reaction volume.
Comparing multiplex systems to singleplex assays can reveal some clear benefits. The requirement for a smaller sample input has already been mentioned; typically, 25–50 L of sample volume are needed to test numerous markers, as opposed to 100 L for each target in an ELISA assay.
Additionally, test dynamic range is frequently improved by multiplex platforms. Multiplex assays are reported to maintain linearity across three or even five orders of magnitude, in contrast to ELISA, which rapidly loses linearity over a few orders of magnitude.
Since targets are frequently present in panels at vastly varied concentrations, linearity over a broad range is crucial for accurate data.
Spatially distinct tests in multiplex immunoassays
The first strategy uses spatially distinct tests, which are like ELISAs but share a single reaction chamber with several antibody pairings. The physical isolation of capture antibodies on a shared solid surface is a characteristic that unites several versions on this topic.
Immobilization of the beads or particles in the multiplex immunoassays
The identification of which analyte is being detected on which bead is a difficulty for the bead immobilisation method for multiplex immunoassays. In order to encode analyte-specific capture beads, methods use mixtures or amounts of dyes.
For instance, Luminex’s method encrypts solid polystyrene microspheres using mixtures of red and near-infrared dyes that may produce codes for 500 different targets. These codes are utilised to identify the captured antibodies that have bound, and phycoerythrin-labeled detector antibodies are employed in a sandwich test to assess protein concentrations.
The first strategy uses spatially distinct tests, which are like ELISAs but share a single reaction chamber with several antibody pairings. The physical isolation of capture antibodies on a shared solid surface is a characteristic that unites several versions on this topic. A second, target-specific detection antibody is employed for measurement once each spot has captured a particular target protein. Spot coordinates can be used as an address to identify the protein being discovered after reading the entire array all at once.
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