What is a multiplex assay?

A multiplex assay is a derivative of ELISA, which can be described as a kind of immunoassay that one can apply through the use of magnetic beads. These magnetic beads can be used to measure several analytes but in just a single experiment.

Concept of a multiplex assay

Multiplex bead array assay is spreading massively and being tagged as a new key discovery, but research shows that it has been in existence for a certain period. Several tests have been carried out through the use of molecular ligands as well as immunological methods.

Advantages of multiplex assays

These multiplex assays have some potentials and benefits that they offer when they are applied, which includes

– Its independent and quantitative ability to assay several analytes simultaneously in meager spaces of the medium used.

– Ability to gather data from various bead arrays to record a statistical figure for the ligands available.

 – It helps reduce the cost of operation, and usage of time is minimized, which stimulates and ensures a timely use in research areas or its related ones.

Application of multiplex assays

To validate assays, some factors such as its precision level, its ability to reproduce, how accurate it is, and how trustworthy it can lead to comparing it with the procedures tagged to be the standards to pass. Almost every array kit comes with a key piece of information and reference that will aid the application. However, there are still certain levels of uncertainty regarding whether the collection of data through multiplex assays will be similar or different from data collected when other methods are applied to get results.

What you need to activate an ELISA assay

The ELISA kit needs some items to be applied before you can be able to use it. There are several of them, and they are so easy to apply.

  • It is important for you to get a microplate reader that can make a correct reading of absorbance at 450nm.
  • You also need to get an automated plate washer, although it can be optional for some operations.
  • Make sure you possess pipettes that tend to precisely dispense 0.5 µl through 1 ml volumes of aqueous solutions.
  • Acquire some multichannel pipettes as they are essential for a very large quantity of samples.
  • Always make sure you have deionized or distilled water in-ground.
  • Have sufficient test tubes in possession for dilution.

What to check out for before selecting ELISA kits for assays

Sensitivity: Sensitivity is a very important factor when choosing your ELISA kit. It is also the lowest detectable amount of a target analyte. ELISA kits ensure a high turnaround time without any compromise on the quality of results.

Stability: ELISA kits of different concentrations get assayed with the same kit for a very long period.

Specificity And Cross Reactivity: Evaluating target analytes for the presence of other proteins may lead to false-positive results. Make sure cross-reactivity is dormant in the kits.

ELISA vs. Multiplex Assays

ELISA, also known as enzyme-linked immunosorbent assay, is the most common assay compared to the multiplex assay. These two have a lot of heavy-weight differences, which distinguishes them from each other.

Differences between Multiplex assays and ELISA

  • Multiplex assay makes use of fluorescence to work out its mode of reporting, while ELISA makes use of an enzyme amplification system of a colorimetric substrate.
  • Multiplex assay seizes its ligands on spherical beads in suspension while ELISA depends on flat surfaces in 96-well plates to capture its own ligands.
  • Multiplex assays are technical and multiplexed which has a potential to subject itself to any change or variation that may come from analyzing a lot of ligands at the same time, out of which an example of such application is cross-reactivity while ELISA only has the ability to generally analyze just an analyte at a time so as to prevent any form of complications coming up from multiplexing.
  • In a multiplex assay system, there is the presence of protein microarray kits that can be applied to capture antibodies in a multiplex fashion.

Comparison between Multiplex Arrays and ELISA

Before a multiplex assay can be used in any clinical research or laboratory process, it has to have a huge potential of achieving similar results to when ELISA was tried out.

Research has indicated that good correlations exist between Multiple assays and ELISA for most of the cytokines tested despite how wide the degree of correlation proved to be. It is probably due to the results obtained when comparisons were made during the assay process.


With various research and studies that have been done between multiplex assays and ELISA measurements for some time, good correlations have been demonstrated, although with the poor occurrence of quantitative values. It has been found out that a comparison of randomly selected multiplex assays and ELISA will most likely result in substantial differences. Multiplex Assays have been established to be easy to perform and can be reproduced. As this multiplex assay is not costly and does not waste time, it will surely continue to be used more and more.