Recently, a new paradigm of xMAP Reader has been provided by Luminex which does not utilize flow cytometry principles for detection. The MAGPIX® Reader System has been developed by Luminex exclusively for MagPlex® microspheres. This technology allows for multiplex analyte determinations on biological samples. By using multiplexed bead sets several analytes can be quantitated simultaneously, saving time, money and precious sample. We are using a panel of known tumor biomarkers to characterize tumor cell lines of known lineage under different conditions to obtain a better understanding of the biology specific to different tumor types.
|Figure 1. Standard Curves for the EMD-Millipore 22-Plex human Circulating Cancer Biomarkers using ELx50 Microplate strip Washer.|
A key element in the ability to multiplex numerous samples is to adequately remove unbound materials while retaining the beads. Here we have used the ELx50 Microplate strip washer to automate the wash steps required to run the 22-Plex human Circulating Biomarkers panel from EMD-Millipore. As demonstrated in Figure 1, individual analyte calibration curves can be generated that will allow concentration determinations for 22 different analytes on each sample.
|Figure 2. HepG2 Cell line Biomarker characterization.|
When dilutions (1:1, 1:3, and 1:6) of conditioned media supernatant from HepG2 cells are analyzed for the secretion of biomarkers a discrete pattern of expression is observed. HepG2 cells demonstrate measurable amounts of AFP, CYFRA21-1, IL-8, MIF, OPN, sFas, and VEGF that correlates with the concentration of the input material.
|Table 1. Determined concentrations of HepG2 cell supernatant|
By, BioTek Instruments