Time-Resolved Fluorescence (TRF) uses fluorescence lifetime to gain information about a molecule or its molecular environment.
As fluorescence’s lifetime is characteristic for each fluorescent molecule and also influenced by the chemical composition of its environment, it can be used to characterize a sample.
When the natural background fluorescence is eliminated, the sample’s fluorescence can be used to deliver highly accurate information about its molecular environment. Aqsens Health’s proprietary invention (E-TRF) enhances this unique capability even further.
The E-TRF fingerprint is formed from a combination of several chemical modulators that have been measured at the same time and in the same sample. The selection of proper modulators is performed using AQH’s proprietary reverse screening process together with our large modulator library.
In E-TRF, each modulator affects the strength of the lanthanide’s luminescence signal in the samples in different ways – some block the luminescence completely, while some modulators have a very minimal effect on the luminescence signal. When the same modulators are used one at a time in different samples together with the lanthanide label and then measured with TRF, the different luminescence signal strengths created by the modulators form a unique luminescence signal profile for each sample – the sample’s fingerprint.
In order to further increase the specificity of the measurements or to classify disease severity, the E-TRF method applies bacteriophages. This is done by modifying the phages’ binding sites to build the phages’ affinity for both dye molecules or quenchers, and biomarkers. This creates a situation in which the color molecules and biomarkers compete over the same binding site. This programming is done through a proprietary biopanning process.
When the phages bind either to the color dye molecule or disease specific biomarkers, the color intensity of the sample changes respectively. These color intensity changes can be measured both with a standard TRF reader or with a smartphone using the AQ MOBI platform and application software. These color reaction changes form the basis for full mobility.
The E-TRF method has several possible applications in screening and healthcare. The agile development framework enables the quick development of accurate and cost-effective screening tests for various different purposes. When developing a new screening test Aqsens Health receives biological samples and the related primary data from clinical research partners. The samples contain control samples and clinically diagnosed disease samples.
Aqsens Health’s modulators and the biological samples are processed in standard 96 microwell plates. The E-TRF measurement process follows the highest quality practices. After the measurement the results are analyzed against the clinical data with the help of statistical application software(ML, AI). When the whole dataset is processed, Aqsens Health has a strong indication of E-TRF’s ability to detect biological disease markers in the sample.