Highly Sensitive & Quantitative Time-Resolved Lateral Flow Immunoassay

6 Davis Drive, Research Triangle Park, NC 27709

Background
BioMedomics team has been involved with nanoparticle technology development for many years. BioMedomics has developed a timeresolved assay platform with its partners and exclusively licensed some key patents associated with time-resolved assay technologies. These technologies include novel long lifetime phosphorescent nanoparticles, nanoparticle conjugation, and time-resolved detection system. In medical diagnostic applications, background interference can severely compromise the sensitivity of fluorescence based detection. Timeresolved fluorescence detection technique has been proven to be capable of providing highly sensitive assays for chemical and biological samples because of its lower background and higher signal/noise ratio than conventional fluorescence technique. However, only a limited number of fluorescence probes have been discovered to possess fluorescence lifetime long enough to significantly differentiate from typical background fluorescence. We developed a new class of phosphorescent nanoparticles with excellent physical and spectral properties for time-resolved biological detection techniques. The particles exhibit strong phosphorescence with long lifetime and large Stoke shift under ambient conditions. There are many potential biomedical applications with such unique nanoparticles. Monodisperse Stable: cross-linked Good flow properties Surface-functionalized Size: 30nm 300nm

Nanoparticles and Results

F N N M N F F N F

Conclusions
BioMedomics has developed its patented point-ofcare (POC) diagnostic platform: Time-Resolved Lateral Flow Immunoassay (TR-LFIA) by applying our proprietary technologies of novel nanoparticles and time-resolved luminescent detection in a lateral flow immunoassay (LFIA). Our nanoparticles have a high loading of the encapsulated phosphorescent molecules to provide strong phosphorescence of long lifetime and high photostability. With our special nanoparticles, lateral flow assays can be very sensitive with the right kinetics of antigen:antibody binding and with the proper selection of suitable reagents and optimization of the running conditions. The TR-LFIA technology has increased test sensitivity by orders of magnitude and provides ultra-low-level quantitation not available in common LFIA tests. As a result, time-resolved detection with its long lifetime probes provides many significant advantages for many clinical applications.

Efficient conversion Quantum Yield ~ 0.2-0.3 Long lifetime: T >100 us Excitation @ 395 nm Emission @ 650 nm

Pt-MTPFPH Pd-MTPFPH
Phosphorescence Emission
Polymer Dye Solvent Encapsulation Crosslinking
1.0
1.0

Phosphorescence Decay

Excitation
0.8
0.8

Phosphorescence
Normalized
Normalized

KC-PP Membrane

0.6 0.4 0.2

0.6 0.4 0.2 0.0

Organic Phosphor material Chloropolymer Matrix Encapsulation Cross-linking Surface-functionalization

Characterization COOH

0.0

Phosphorescent particle preparation process

350

400

450

500

550

600

650

700

0.0

0.1

0.2

0.3 Time (us)

0.4

0.5

0.6

Future Directions
The novel nanopartilcles developed should find a wide variety of applications for detection of chemical and biological molecules. Based on our novel nanotechnologies and other capabilities, BioMedomics will work with strategic partners to co-develop diagnostic tests of specific interest to them. We are particularly interested to work on early cancer diagnosis with unique biomarkers, heart diseases diagnosis, as well as infectious disease detection.

Wavelength (nm)

Application to Quantitative Point-of-Care Diagnostics

Sample Conjugate: Binder I-Probe Calibration Line

High Intensity Long lifetime Targeted excitation/emission

Time-Resolved Bioassays
The time-resolved bioassays separate the fluorescence of interest from the background through lifetime differences. The technique involves exciting a fluorescent label of a long lifetime with a short pulse of light, then waiting a period of time for the background and other unwanted fluorescence to decay to a low level before collecting the remaining long-lived fluorescent signal. The ability to eliminate the background is especially important for biomedical diagnostics. Time-resolved methods have been reported to increase the detection sensitivity by two or more orders of magnitudes over conventional fluorescence detection.
Detection Line: Binder II

Application for C-Reactive Protein Detection

4
0.03

3
0.02

Signal

2 1

Signal

0.01

Background and Interference

40

0.00

0
Blood Fluorescence

500

1000 1500 2000 2500 3000 3500

10
Pt-KCP (ng)

15

20

Pt-KCP (ng)

Arbitrary Unit

20
Membrane

Contact:
Dr. Frank Wang, President & CEO, fwang@biomedomics.com, (919) 883-5542 David Hocking, VP of Business Development, dhocking@biomedomics.com, (919) 558-1228.

Time-Resolved Signal

Background-corrected signal

0 500 550 600 650 700

Detection Sensitivity: <0.2pg/ml

Wavelength (nm)