Sensor system for saccharides

• US 7,358,094 B2
• Filed: 05/01/2003
• Issued: 04/15/2008
• Est. Priority Date: 05/01/2003
• Status: Active Grant

• Current Assignee: Baxter Healthcare SA (Baxter International Inc.), Baxter International Inc.
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: James, Tony D., Arimori, Susumu, Bell, Michael L.
• Primary Examiner(s): Warden; Jill
• Assistant Examiner(s): TURK, NEIL N
• Application Number:
  US10/427,404
• Publication Number:
  US 20040219535A1
• Time in Patent Office: 1,811 Days
• Field of Search: 435/14, 436/14, 436/94, 422/50
• US Class Current: 436/172
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

An analytical system for detecting a saccharide is provided. The analytical system includes a saccharide sensor attached to a solid substrate and having the formula:

Where Z is selected from the group consisting of hydrogen, alkyl groups, an aryl; N is a nitrogen atom; B_d1 and B_d2 are independently selected binding groups, wherein the binding groups are capable of binding an saccharide; Sp is a spacer; An is an anchor group for attaching the sensor to solid substrate, including a bead; and n=1 or 2, m=1 or 2, y=1 or 2, and p is an integer. In one embodiment the saccharide sensor preferentially detects glucose. The invention further provides beads having the saccharide sensor attached thereto and methods of detecting glucose using the system.

Claims

1. An analytical system for detecting a saccharide comprising:

a) a sample known to contain or suspected of containing a saccharide;

b) one or more than one solid substrate, the solid substrate having attached thereto one or more than one saccharide sensor, the sensor having the formula;

2. The system of claim 1 further comprising a labeled glucose analogue that competes with glucose for binding with the saccharide sensor.

3. The system of claim 1, wherein the solid substrate is one or more than one bead having identification labels comprising:

a) a fluorescent analyte detection dye, the analyte detection dye being capable of being excited by light at a first excitation wavelength and capable of emitting light at a peak wavelength when excited, and

b) two or more than two fluorescent labels in a first combination of relative amounts, the fluorescent labels being capable of being excited by light of a same second excitation wavelength and capable of emitting lights at first and second peak wavelengths, distinguishable from each other, respectively, wherein the peak wavelength of the emitted light of the analyte detection dye is different from the first and second maximum wavelengths of the emitted lights of the fluorescent labels by at least 100 nm, and the first and second peak wavelengths differ by at least 100 nm and one of the excitation wavelengths is greater than about 750 nm.

4. The system of claim 3 wherein the means for detecting the particle and saccharide in the sample comprises:

a) means for exciting the fluorescent dye;

b) means for exciting the first and second fluorescent labels;

c) means for detecting the emitted lights; and

d) means for correlating the detected emitted lights with a particular bead under analysis.

5. The system of claim 3 wherein the analyte detection dye is externally complexed to the outside of the bead, and the fluorescent labels are embedded within the bead.

6. The system of claim 3 wherein the fluorescent labels are both cyanine dyes having emitting lights greater than 750 nm.

7. The system of claim 3 wherein light at the first excitation wavelength causes substantially no emitted light by the fluorescent labels and light at the second excitation wavelength causes substantially no emitted light by the analyte detection dye.

8. A saccharide sensing substrate, comprising

a solid substrate;

and

a saccharide sensor attached to the substrate having the formula;

9. The saccharide sensing substrate of claim 8, wherein B_d1 is R₁—

B(OH)₂ and B₂ is R₂—

B(OH)₂, B_d1 and B₂ are capable of binding a saccharide molecule to form a stable 1;

1 complex, and R₁ and R₂ are aliphatic or aromatic functional groups selected independently from each other and B is a boron atom.

10. The saccharide sensing substrate of claim 9, wherein R₁ and R₂ are selected from the group consisting of:

methyl, ethyl, propyl, butyl, phenyl, methoxy, ethoxy, butoxy, and phenoxy groups.

11. The saccharide sensing substrate of claim 8, wherein Sp is a straight-chain alkane.

12. The saccharide sensing substrate of claim 11, wherein the straight-chain alkane comprises from 1 to 9 carbon atoms.

13. The saccharide sensing substrate of claim 12, wherein the straight-chain alkane comprises 6 carbon atoms.

14. The saccharide sensing substrate of claim 8, wherein the saccharide sensor attached to the solid substrate has the following general structure:

15. The saccharide sensing substrate of claim 8, wherein the sensor attached to the solid substrate has the following general structure:

16. The saccharide sensing substrate of claim 8, wherein B_d1, B_d2, Sp, m, and q are chosen to provide selective binding of the saccharide sensor to glucose.

17. The substrate of claim 8, wherein the saccharide sensor is a glucose binding sensor and the saccharide sensing substrate has the formula:

18. An analytical system for detecting a saccharide comprising:

a) a sample known to contain or suspected of containing a saccharide;

b) one or more than one solid substrate, the solid substrate having attached thereto one or more than one saccharide sensor having the formula;

19. The analytical system of claim 18, wherein Z is selected from the group consisting of hydrogen, alkyl groups and a phenyl group.

20. The analytical system of claim 19, the system further comprising a labeled glucose analog that competes with glucose for binding with the saccharide sensor.

21. The system of claim 20 wherein the labeled glucose analog is a propane diol ring-locked cyanine dye having the formula:

22. The system of claim 18, wherein Z is a fluorophore.