TR-FRET immunoassay is an analytical method which can be used for the detection and quantification of specific proteins, biomolecules, peptides, cytokines, hormones, antibodies, etc present in serum, plasma and cell culture supernatant. This method is fast, wash-free, eliminates background fluorescence and economical.
Our Bright-Dtech™ technology brings a more sensible (high brightness resulting in an improved limit of detection) and stable (high photo-stability over days or weeks, resistance to photo-bleaching and durable signal intensity) approach.
In Bright-Dtech™ TR-FRET method, two antibodies are used to detect the protein of interest. The first antibody is conjugated to our fluorescent nanoparticles (donor) and the second one is coupled with fluorescent molecules (acceptor). If both antibodies recognize the target, the fluorescent intensity of the donor is transferred to the acceptor and the signal is detected using time-resolved fluorescence reader to obtain a specific and quantitative result. More informations about TRF and TR-FRET in the Technology page (Click on the below buttom).
Required material : Black microplates, plate shaker, Time-Resolved Fluorescence microplate reader.
Why using Bright-Dtech™ for TR-FRET applications?
Our technology brings benefits to users. Thanks to Bright-Dtech™ TR-FRET there is no substrate needed. We have a very low background, an exceptional lifetime and no photobleaching. We have also a low LOD and LOQ, a low hands-on time, no-wash and easy to implement. Moreover, our nanoparticles are compatible with various acceptors.
Example: Immunoassay in TR-FRET assessing IgG antibodies
Samples or standards and detection reagents are added to microtiter plate wells. After incubation, Time-Resolved Fluorescence (FRET) is measured at a particular wavelength. The intensity of the resulting signal is directly proportional to the concentration of the antigen present in the sample.
We have shown that Bright-Dtech™ TR-FRET technology is able to significantly enhance the IgG antibody detection thanks to the exceptional brightness of our fluorescent nanoparticles.
Figure 1. TR-FRET measurements for human IgG antibody detection.
Bright-Dtech™ technology enhances the detection of human IgG antibodies due to its high specificity and sensitivity given by the time-resolved fluorescence signal in FRET. The assay range was 1-10 000 pg/mL leading to a very low/significant limit of detection around 512 pg/mL (using 3 standard deviation).