On this episode of Molecular Moments, Chad Briscoe speaks with Dr. David Berkowitz, Director of the Division of Chemistry at
BioAgilytix employs Bio-Rad’s Luminex-based Bio-Plex Multiplex System with laser-based technology and Luminex FLEXMAP 3D® systems to run bead-based immunoassays. These platforms give us the ability to multiplex many different assays simultaneously, providing greater information from less sample volume and in a shorter timeframe than required for traditional immunoassay methods.
Based on the principles of flow cytometry, the Bio-Plex platform utilizes xMAP technology licensed from Luminex to deliver its multiplexed assay capabilities. This analyzer combines advanced fluidics, optics, and digital signal processing with Luminex’s proprietary technology to perform discrete assays on the surface of color-coded beads known as ‘microspheres’.
Built around three core technologies – the fluorescently dyed microspheres, or beads, provided from Luminex; a flow cytometer with two lasers and associated optics; and a high-speed digital processor able to efficiently manage digital output – the platform employs fluorescent methods for detection, using the bead sets to identify the analytes, and detection antibodies to measure the quantity of analyte. The use of different colored beads allows for simultaneous detection of many analytes in the same sample, and laser excitation is employed to determine analyte concentration.
The main advantages of the Luminex FLEXMAP 3D system are the ability to multiplex up to 500 analytes with minimal sample volume, and automation to support high-throughput bioassay applications.
Luminex is ideal for bioanalytical applications that require both high plex and high throughput at the same time. It is able to yield dramatically more information from rare and / or limited samples, and decipher complex interrelationships among proteins. While traditional ELISA and other technologies excel at high throughput applications – greater than 1,000 samples per day – they lack the ability to multiplex more than five tests at a time. Alternatively, microarray technology can perform high density screening – sometimes more than 250-plex tests – but cannot yield the reproducibility needed for high throughput applications.
As a flow-based system, our Luminex platforms read and quantitate bead events individually, and can perform many assays on each sample. For example, a 30-plex kit can obtain up to 3,000 data points a day on up to 100 samples. BioAgilytix most frequently uses Luminex in our work with biomarkers and cell-based assays. Our team has optimized and validated several cytokine, chemokine, growth factor, and biomarker assays ranging from known vendor protein panels (i.e., pro- or anti-inflammatory panels) to completely customized panels. We can also perform non-regulated analysis of antibodies to specific HLA haplotype expressed by allogenous cell therapy using Luminex.