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Simultaneous Quantitation Of Cytokines, Chemokines, Growth Factors Using Luminex ELISA Assay

Luminex multiplex assays are specifically designed to facilitate the detection and quantification of multiple analytes such as chemokines, cytokines, growth factors, and expressed genes. Luminex-based assays produce results similar to conventional ELISA and qPCR assays but are more efficient and provide greater throughput. Luminex cytokine and chemokine assays combine advanced optics, digital signal processing, and fluidics with microsphere technology to provide multiplexed capabilities. Luminex platforms are flexible and open to configure into a wide array of bioassays as quickly and cost-effectively as possible.

The Luminex platform uses superparamagnetic microspheres and a polystyrene surface. The microspheres are internally dyed with different fractions of red and infrared fluorophores that can generate 100 unique spectral signatures. Each unique signature corresponds to a distinct bead area. Conjugating a distinct bead with a distinct antibody helps to identify multiple target proteins in a single well. Multiple protein-specific beads are then combined in a single well and facilitate simultaneous detection and quantification of several proteins in a single well.

Types of Luminex platforms

Luminex provides four different instrument platforms; Luminex 200, INTELLIFLEX, FLEXMAP 3D, and MAGPIX. Each of these instruments has analyzers either based on the principles of quantitative fluorescent microscopy or fluorescent flow cytometry. However, MAGPIX has now been discontinued.

Luminex 200 are industry-standard instruments that can conduct up to 80 types of tests in a single assay volume. It is based on the principle of flow cytometry. INTELLIFLEX is the latest edition in the Luminex family. They can conduct up to 80 tests and are specifically built for high throughput. INTELLIFLEX is equipped with upgraded hardware, a side eject, and a functional dual reporter. Similarly, FLEXMAP 3D facilitates up to 80 different tests and is compatible with 96 and 384-well plates.

Specialized Luminex ELISA validation assay.

The Cytokine/Chemokine/Growth Factor 45-Plex ProcartaPlex Panel 1 is a specialized Luminex-based multiplex assay kit capable of analyzing 45 analytes in a single well. Such multiplexing capabilities help explore the underlying principles of the immune system. The system comprises five sub-panels enabling a broader screening of the target proteins and subsequent analysis of specific subsets of cytokines that can be added separately. The sub-panels can be combined with each other or used individually in simplex formats.

The panels are extensively tested for cross-reactivity, interference, and analyte combinability. The multiplex kit is available for six species including, human, nonhuman primates, rats, mice, canine and porcine. ProcartaPlex assays can provide results in just four hours and require as little as 50 µL cell culture samples or 25 µL serum or plasma samples. Moreover, its multiplexing capabilities can help detect and quantify up to 65 analytes of interest in a single assay sample.

The conclusion

Luminex-based assays are the primary choice of analytical tool in the detection and quantification of cytokines, chemokines, and growth factors in biological samples. With a wide variety of assays available and the option of custom-made assays, researchers have numerous assay kits available in studying and evaluating the immune system.