Luminex Style Assays using the Bioplex
Bio-plex applications available in APAF
Bio-Plex/Luminex systems allows for the simultaneous analysis of multiple analytes in samples - including cytokines, chemokines, growth factors and phosphoproteins. Commercial kits are also available for various assays such as pro-acute phase proteins, pro-angiogenesis, pro-diabetes, cell signaling, neuroscience, bone metabolism, iso-typing and cancer biomarkers that can be analyzed with Bio-Plex/Luminex systems.
The principle of the assay is similar to a capture sandwich immunoassay - an antibody to the target protein including very low abundant proteins that are difficult to identify and quantify with other proteomic technologies is covalently coupled to internally dyed beads. When the beads are incubated with sample, the protein of interest is captured and then a biotinylated antibody for a different epitope is added to the reaction, which is then detected with streptavidin-phycoerythrin. Using a dual-laser flow-based reader, beads are analyzed for the detection antibody and the internal bead signature, identifying both the protein analyzed and the level bound to the bead. Measuring 42 analytes (42-plex kit) in 40 samples in duplicate with Bio-Plex/Luminex system takes only a couple of hours.
Key benefits of multiplexed assays
- Significantly decreases experimental variables - multiplexing up to 100 bioassays per well (per sample).
- Significantly increases the amount of information from a single sample - using 50 µL of sample obtains up to 100 results (target peptides and proteins).
- Quantitative results - over a 3 to 4 log dynamic range, with strong concordance to ELISA and Mass Spectrometry.
- Automated liquid handling - All the liquid handling operations for cytokine assays performed on a robotic motion system for precise and reproducible liquid delivery in an automatic fashion.
- Cost effective – reduces reagent and labor cost.
APAF offers analytical services to clients for use of cell culture media, serum/plasma or other biofluids and tissues samples (human and rodent). Dedicated robotic liquid handling enables reproducible assays.
Dr. Alamgir Khan