MSD Assay Details
The Meso Scale Discovery platform is the gold standard for multiplex immunoassays. Here is why:
- Multiplexing allows fast and cost-efficient testing of an extensive array of cytokines/chemokines
- A small sample volume of as little as 5 µl is sufficient to enable testing of plasma from interim blood draws
- The exceptionally broad dynamic range enables the detection of proteins that are present in abundance as well as in tiny amounts (up to sub-fg/ml, which is 1000 times more sensitive than standard ELISA)
- Meso Scale Discovery provides ready-to-use kits which are thoroughly validated and include defined calibrators for reliable data with high reproducibility
MSD assays at Reaction Biology are performed with 96 well plates containing up to 10 detection spots per well.
High binding carbon electrodes on the bottom of the plates allow for attachment of capture antibodies with a 10x greater binding capacity than commonly used plates.
Most MSD immunoassays are sandwich assays similar to ELISA with the detection antibody labeled with a SULFO-tag. During readout, electricity is applied to the plate electrodes by an MSD instrument leading to light emission by the SULFO-tags. The light is then measured to quantify analytes in the sample.
Example study: Quantification of recombinant antibodies from blood samples
Mice were treated at day 0 with a viral construct for expression of a recombinant antibody.
Blood samples (about 20 µl) were taken at the indicated time points, starting with a first sample one day before application of the virus in two different doses.
The resulting expression of a recombinant antibody in mouse serum was determined with MSD technology using antigen-coated assay plates and quantified against an antibody standard (graph below).
Example study: Testing the effect of different cell culture conditions on cytokine release.
Levels of cytokines IL-1beta and IL-8 were analyzed via MSD technology in cell culture supernatants of human mesenchymal stem cells cultured in different conditions for 1, 3, or 7 days. Cytokines were measured in multiplex fashion and quantified against a recombinant calibrator (lower graphs). N =3
Instrument: MSD MESO QuickPlex SQ 120
Selection of analytes:
- Cytokines and Chemokines
- Intracellular signaling assays including GPCR ligand-binding proteins, phosphoproteins, and ubiquitinated proteins
- Metabolic assays
- Oncology biomarkers
- Toxicology markers
Please reach out to have a conversation with our scientists about the setup best suited to answer your specific research questions.
- Plasma or other body fluids
- Cell culture supernatant
- Tissue homogenates
- Cell lysates
Please discuss with us how we can help to derive or prepare the samples for your study.
For maximum sensitivity, undiluted samples of at least 25 µl are required. However, the high sensitivity of MSD assays allows for most targets to predilute the sample enabling us to perform multiplex analysis out of tiny sample volumes, such as frequent bleeds of mice. Here we can work with a sample volume of as little as 5 µl.