Development and validation of home-made Luminex assays for the quantification of multiple cytokines in human samples

Cytokine profiling is a powerful tool to link the host immune system with disease pathogenesis and/or treatment efficacy. The analysis of cytokine expression is frequently required for the characterisation of the status of the immune system, to demonstrate the functionality of the cells of interest and as an analysis of surrogate markers for in vitro potency testing.

The aim of this study was to design a Luminex immunoassay using home-made coupled magnetic beads with specific monoclonal antibody for the quantification of human TNFα, IL-2 and Perforin, on the Luminex MAGPIX analyser. The performance of this 3-plex assay was evaluated in terms of accuracy, precision, quantification range and sensitivity.

This study demonstrates the successful development and validation of home-made microbead-based Luminex assay for the quantification of 3 cytokines, and the ability of these home-made coupled beads to be combined with a commercial multiplex assay.

It also demonstrates the ability to effectively couple custom microbeads with capture antibodies which allows for the development of custom multiplex assays with equivalent performance compared to commercial kits.