Fiber-optic biosensor and biosensing methods

• US 8,563,328 B2
• Filed: 02/16/2006
• Issued: 10/22/2013
• Est. Priority Date: 07/26/2004
• Status: Expired due to Fees

• Current Assignee: Research Foundation Incorporated University of Louisville, University of Cincinnati
• Original Assignee: University of Louisville
• Inventors: Kang, Kyung Aih, Ahn, Chong H.
• Primary Examiner(s): Chin, Chris L
• Application Number:
  US11/355,467
• Publication Number:
  US 20060286680A1
• Time in Patent Office: 2,805 Days
• Field of Search: 435/7.1, 435/7.94, 436/172
• US Class Current: 436/518
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

A method for biosensing that includes passing, via convective flow, a sample believed to contain one or more target biomarkers through a microfluidic channel and over the surface of an optical waveguide that has been prepared to bind the one or more target biomarkers, and sensing for an emission output from the optical waveguide at a wavelength that is characteristic of the binding of the target biomarker. A biosensor device that includes a module defining at least one microfluidic channel, an optical waveguide exposed along at least a portion of its length to fluid flow within the microfluidic channel, where a surface of the optical waveguide being prepared to bind a target biomarker, and an excitation source to couple an excitation wavelength of light into the optical waveguide. The device also includes a sensor for detecting emission light from the optical waveguide at an emission wavelength characteristic of binding of the target biomarker.

Claims

1. A biosensing method for simultaneously detecting and quantifying two or more target biomarkers, said method comprising:

providing at least two optical waveguides, each of the at least two optical waveguides being disposed at least partially in a respective microfluidic channel of a serpentine shaped chamber, the at least two optical waveguides being fluidly coupled to one another in series within the serpentine shaped chamber;

selecting 1°

Mabs corresponding to each of the two or more target biomarkers;

immobilizing the 1°

Mabs on the optical waveguides;

exposing to the 1°

Mabs a sample believed to contain the target biomarkers using convective flow within the serpentine shaped chamber, wherein the sample flows through the serpentine shaped chamber and flows over each of the at least two optical waveguides in series;

providing fluorophore-tagged 2°

Mabs corresponding to the target biomarkers;

exciting the at least two optical waveguides; and

observing a fluorescence emission to determine the presence and quantity of the target biomarkers.

2. The method of claim 1 wherein said providing at least two optical waveguides comprises providing at least two quartz optical sensing fibers coupled to one another in series.

3. The method of claim 1 wherein said providing at least two optical waveguides comprises providing four optical sensing fibers coupled to one another in series.

4. The method of claim 3 further comprising providing four 1°

Mabs corresponding respectively to Cardiac troponin I (cTnI), myoglobin (MG), B-type natriuretic peptide (BNP) and C-reactive protein (CCRP).

5. The method of claim 3 further comprising providing four 1°

Mabs corresponding respectively to PC, PS, ATIII and PLG.

6. The method of claim 1 wherein said selecting 1°

Mabs comprises selecting the 1° and

2°

Mabs such that the 1°

has lower affinity with respect to the target biomarker than does the 2°

Mab.

7. The method of claim 1 wherein said providing at least two optical waveguides comprises providing at least two optical sensing fibers coupled to one another in series, and wherein said immobilizing the 1°

Mabs on the optical waveguides comprises immobilizing the 1°

Mab via an avidin-biotin bridge on a surface of the optical sensing fibers.

8. The method of claim 1 further comprising providing fluorescence enhancers to enhance fluorescence emission of the fluorophore.

9. The method of claim 8 wherein said providing fluorescence enhancers comprises providing a nanometal particle at a predetermined distance from the fluorophore.

10. The method of claim 9 wherein said providing a nanometal particle comprises providing one of the group consisting of nanogold particles, nanosilver particles, a reagent containing nanogold particles and a reagent containing nanosilver particles.

11. The method of claim 9 wherein said providing fluorescence enhancers further comprises surrounding the nanometal particle with a self-assembled monolayer to maintain the predetermined distance between the fluorophore and the nanometal particle;

wherein the nanometal particle is dispersed within a solution.

12. The method of claim 1 wherein said providing at least two optical waveguides comprises providing at least two optical sensor fibers coupled to one another in series, and wherein said immobilizing the 1°

Mabs on the optical waveguide comprises regenerating the optical sensing fibers.

13. The method of claim 1 wherein said providing at least two optical waveguides comprises providing at least two optical sensor fibers coupled to one another in series, and wherein said immobilizing the 1°

Mabs on the optical waveguide comprises coupling a fluorometer to the optical sensing fibers to determine fluorescence emission.

14. The method of claim 1 further comprising selecting the fluorophore from the group consisting of Cyanine 5, AF647, ATTO 647, Bodipy 650/665 and DY-636.

15. The biosensing method of claim 1 further comprising providing a serpentine structure within said serpentine shaped chamber to enhance molecular binding efficiency.

16. The biosensing method of claim 1 wherein the serpentine shaped chamber comprises a bump structure disposed at inner surfaces of the microfluidic channels within the serpentine shaped chamber.

17. The biosensing method of claim 1, wherein the respective microfluidic channels are disposed in parallel to one another and fluidly connected to one another via communication channels.

18. The biosensing method of claim 1, wherein the serpentine chamber has an inlet and an outlet, and wherein said exposing further comprises the sample flowing in sequence through the inlet, over each of the at least two optical waveguides in series, and then through the outlet.