Sensor system for saccharides

• US 20040219535A1
• Filed: 05/01/2003
• Published: 11/04/2004
• 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: Arimori, Susumu, Bell, Michael L., James, Tony D.
• Granted Patent: US 7,358,094 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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; S is the 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 having the formula;

wherein;

Z is selected from the group consisting of hydrogen, alkyl groups, and aryl groups;

B_d1 and B_d2 are independently selected binding groups;

N is a nitrogen atom;

Sp is an aliphatic spacer;

An is an anchor group for attaching the sensor to the solid substrate;

S is a solid substrate; and

n=1 or 2, m=1 or 2, y=1 or 2, and x is an integer; and

c) means for detecting at least one of saccharide that is bound to the sensor or saccharide that is not bound to the sensor.

2. The sensor of claim 1, wherein B_d1 is R₁—

B(OH)₂ and B_d2 is R₂—

B(OH)₂, B_d1 and B_d2 are capable of binding a saccharride 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.

3. The system of claim 2, wherein R₁ and R₂ are selected from the group consisting of:

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

4. The system of claim 1, wherein Sp is a straight-chain alkane.

5. The system of claim 4, wherein the straight-chain alkane comprises from 1 to 9 carbon atoms.

6. The system of claim 5, wherein the straight-chain alkane comprises 6 carbon atoms.

7. The system of claim 1, wherein the saccharide sensor attached to the solid substrate has the following general structure:

8. The system of claim 1, wherein the sensor attached to the solid substrate has the following general structure:

9. The system of claim 1, wherein B_d1, B_d2, Sp, m, and q are chosen to provide selective binding of the saccharide sensor to glucose.

10. The system of claim 9 further comprising a labeled glucose analogue that competes with glucose for binding with the saccharide sensor.

11. 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.

12. The system of claim 11 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.

13. The system of claim 11 wherein the analyte detection dye is externally complexed to the outside of the bead, and the fluorescent labels are embedded within the bead.

14. The system of claim 11 wherein the fluorescent labels are both cyanine dyes having emitting lights greater than 750 nm.

15. The system of claim 11 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.

18. A method of detecting a saccharide comprising:

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

b) providing an analytical system according to claim 1;

and

c) detecting a saccharide in the sample.

19. The method for detecting an saccharide of claim 18 wherein the saccharide in the sample that is detected is glucose.

20. The method for detecting an analyte of claim 19 wherein the analytical system further comprises a labeled glucose analogue that competes with glucose for binding with the saccharide sensor and glucose is detected by a competitive assay.

16. A saccharide sensing bead, the bead having a saccharide sensor attached to it having the formula:

17. The bead of claim 16, wherein the saccharide sensing bead has the formula: