Overview
This invention describes an improved method for conducting a bioassay using fluorescence intensity or lifetime fluorescent emissions. This new method avoids the need of complex enzymatic and biochemical amplification of the fluorescence signal. Although it has applicability over a wide range of bioassays, the method has particular application in sensing, imaging and assaying cytokines. MEF-spot allows for the rapid detection of cytokines and other proteins secreted by a living cell in small quantities, in the presence of living cells and without directly rinsing the sample before conducting the bioassay. In addition to imaging individual cell secretions, this approach allows the visualization of cell-cell interactions in co-cultured cells.
The sensing and imaging principles of MEFspot rely on the enhancement of fluorescence intensity or changes in fluorescence decay of an analyte bound to a surface with a fluorophore-labeled detection probe. The sensing bioactive surface comprises the capture biomolecule attached to the surface with plasmonic nanostructures. This technology also includes the fabrication procedure of plasmonic substrates. The binding of the detection probe to the bioactive surface with an analyte increases fluorescence intensity up to 200x and a decrease in fluorescence lifetime by more than 5x. The enhancement of fluorescence intensity allows for direct sensing and imaging of biomolecule interactions.
MEFspot is a significant advance in single cell assays as its sensitivity is comparable to enzyme-linked immunospot but it uses a substantially simpler procedure that still allows for real-time monitoring and quantitative analysis.
Applications
Current techniques for secreted cytokine assays include ELISA, flow cytometry, RT-PCR, ELISPOT, and FLUOROSPOT. End users for these bioassays include research labs, hospitals, blood banks, CROs, biotech companies, and clinical labs. MEFspot has the potential to be used as a life science reagent, in clinical diagnostics and therapeutics, and in drug development. In 2016, the global immunoassay market reached $16 billion at a compound annual growth rate of 8.2%. It is expected to reach $25 billion by 2021.
Advantages
Does not require enzymatic or biochemical amplification
Assay can be performed in the presence of living cells and other analytes
Cytokine detection of ~ 0.01 ppb
Can assay cell-cell interactions in real time
Stage of Development
The technology has been extensively researched by the inventors. Peer review articles demonstrating the technique on numerous analytes have been published.
(As of 5/17/2017)- MEW
Licensing Potential
Available for licensing and commercial development
Contact Info
Office of Technology Transfer
620 W Lexington St., 4th Floor
Baltimore, MD 21201
Email: [email protected]
Phone: (410) 706-2380