Glucose biosensor

• US 6,475,750 B1
• Filed: 08/23/2000
• Issued: 11/05/2002
• Est. Priority Date: 05/11/1999
• Status: Expired due to Fees

• Current Assignee: M-Biotech, Inc.
• Original Assignee: M-Biotech, Inc.
• Inventors: Bae, You Han, Jung, Dal Young, Han, In Suk, Magda, Jules John
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/644,323
• Time in Patent Office: 804 Days
• Field of Search: 435/14, 435/25, 435/26, 435/287.1, 436/95, 436/148, 604/891.1, 604/892.1
• US Class Current: 435/14
• CPC Class Codes: A61B 5/0002 Remote monitoring of patients using telemetry, e.g. transmission of vital signals via a communication network A61B5/07 takes precedence; transmission systems for measured values G08C; transmission H04B; transmission of digital information H04L; wireless communication networks H04W
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/006 for glucose
• CPC Class Codes: G01N 13/04 Investigating osmotic effects
• CPC Class Codes: G01N 7/00 Analysing materials by measuring the pressure or volume of a gas or vapour
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

A biosensor (10) has a hydrogel (30) in a rigid and referably biocompatible enclosure (20). The hydrogel (30) includes an immobilized glucose-binding molecule such as concanavalin A (Con A) and an immobilized hexose saccharide such as a-D-mannopyranoside. The immobilized hexose saccharide competitively binds with free glucose to the glucose-binding molecules, thus changing the number of crosslinks in the hydrogel (30), which changes hydrogel swelling tendency and the pressure of the hydrogel in its confined space in proportion to the concentration of free glucose. By measuring the change in hydrogel pressure with a pressure transducer (40), the biosensor (10) is able to accurately measure the concentration of the tree glucose molecule without the problem of oxygen limitations and interference encountered by prior art biosensors. A battery (64) powered telemeter (60) operably engaged to the pressure transducer (40) sends a radio data signal to a receiver (66) containing an alarm system operably attached to a computer (62). Furthermore, an alarm system utilizes such a sensor to automatically notify a person that the blood glucose level is outside a predetermined parameter, and/or to automatically inject an agent such as glucose or glycogen which will raise blood glucose levels.

Claims

1. A biosensor for measuring the concentration of glucose, the biosensor comprising:

a polymeric hydrogel that changes its crosslinking density and swelling tendency in proportion to the concentration of free glucose;

glucose binding molecules chemically or physically immobilized in the hydrogel;

a hexose saccharide or a polysaccharide chemically or physically immobilized in the hydrogel, the free glucose competitively binds with the immobilized hexose saccharide or polysaccharide to the glucose binding molecules, which reduces the number of hydrogel crosslinks, thereby causing the hydrogel to swell and increasing the pressure exerted by the hydrogel in its enclosure;

a means for measuring the pressure or amount of swelling of the hydrogel; and

a means for reporting the concentration of the glucose molecule based on the measured pressure of the hydrogel.

2. The biosensor of claim 1 further comprising a rigid enclosure containing the hydrogel, the enclosure having an open end sealed by a semipermeable membrane that allows glucose molecule to diffuse into the hydrogel.

3. The biosensor of claim 2 wherein the enclosure further includes a diaphragm, the hydrogel being enclosed between the diaphragm and the semipermeable membrane, the diaphragm working in conjunction with the means for measuring in order to monitor changes in the pressure and swelling tendency of the hydrogel.

4. The biosensor of claim 3 wherein the enclosure is conjugated with heparin and polyethylene glycol.

5. The biosensor of claim 3 wherein the enclosure is coated with a semipermeable membrane and a biodegradable polymer on the semipermeable membrane.

6. The biosensor of claim 5 wherein the biodegradable polymer is conjugated with heparin and polyethylene glycol.

7. The biosensor of claim 5 wherein the semipermeable membrane has a pore size between 0.1 to 15 μ

m.

8. The biosensor of claim 1 wherein the hydrogel includes crosslinking that allows small molecules such as glucose to diffuse into the hydrogel, but prevents diffusion of any macromolecules such as glycoproteins, lectins, and polysaccharides.

9. The biosensor of claim 1 wherein the biosensor includes a hexose saccharide with a vinyl group conjugated to the C1 hydroxyl group of the hexose saccharide, the vinyl group being used to chemically immobilize the hexose saccharide to the polymer backbone of the hydrogel.

10. The biosensor of claim 9, wherein the hexose saccharide is selected from the group that consists of α

-D-mannopyranoside, p-nitrophenyl-α

-D-mannopyranoside, or p-nitrophenyl-α

-D-glucopyranoside.

11. The biosensor of claim 1 wherein the hexose saccharide includes a monomer containing hexose moieties such as glycidyl acrylate, glycidyl butyl ether, glycidyl cinnamate, or glycidyl methacylate such as glycosyloxyethyl methacrylate.

12. The biosensor of claim 1 wherein the polysaccharide chemically or physically immobilized in the hydrogel is any macromolecule that contains polysaccharides, such as for example glycogen, starch, cellulose, nigeran, paramylon, luteose, mucin, and glycoproteins.

13. The biosensor of claim 1 wherein the glucose binding molecules are chosen from the group consisting of glucose oxidase, hexokinase, glucosidase, xylose. isomerase, glucose phosphorylase (glucokinase), lactate dehydrogenase, boronic acids, and lectins.

14. The biosensor of claim 13 wherein the glucose binding molecules include genetically modified proteins, which have only binding sites to glucose moieties but having no enzymatic activity.

15. The biosensor of claim 1 wherein the hydrogel is biocompatible, nontoxic, and inert in the body.

16. The biosensor of claim 1 wherein the hydrogel has porous structure to enhance diffusion of glucose into the hydrogel.

17. The biosensor of claim 1 wherein the hydrogel further includes chemically immobilized pendant groups (pKa 3-11) which are charged under physiological pH conditions, with the density of charged pendant groups chosen to optimize the amount of hydrogel swelling in response to free glucose concentration changes.

18. The biosensor of claim 1 wherein the means for measuring hydrogel pressure or swelling tendency is a pressure transducer.

19. The biosensor of claim 1 wherein the means for measuring hydrogel pressure or swelling tendency is a piezoelectric transducer or a piezoresistive pressure sensor.

20. The biosensor of claim 1 wherein the means for reporting is a battery powered telemeter that transmits a data signal to a receiver operably connected to a computer means, the computer means comparing the data signal to a calibration curve to calculate the concentration of glucose, the computer means then reporting the concentration through a reporting means.

21. The biosensor of claim 20 wherein the means for reporting is a computer electrically connected to the means for measuring, the computer comparing data from the means for measuring to a calibration curve to calculate the concentration of the glucose molecule, the computer means then reporting the concentration through a reporting means.

26. The biosensor of claim 1, wherein the hexose saccharide is chosen from the group of hexose saccharides consisting of glucose and mannose.

27. The biosensor of claim 1, wherein the polysaccharide is chosen from the group of polysaccharides consisting of mucin, glycogen, and glycoproteins.

28. The biosensor of claim 13, wherein the lectins are chosen from the group of Con A and isolectin I.

29. The biosensor of claim 1, additionally including means for generating an alarm signal responsive to the reported concentration of the glucose if outside a predetermined range, and a transmitter for transmitting the alarm signal to a remote location.

30. The biosensor of claim 29, additionally including a receiver for receiving alarm signals from the transmitter, and a dialer for dialing a telephone in response to an alarm signal received from the transmitter.

31. The biosensor of claim 29, additionally including an injection device for injecting agents into a person in response to alarm signals from the means for generating an alarm signal.

32. The biosensor of claim 1, additionally including an injection device for injecting agents into a person in response to reported concentration of the glucose.

22. A biosensor for measuring the concentration of glucose in a person, the biosensor comprising:

a rigid, biocompatible enclosure having an open end and a closed end, the open end being covered by a semipermeable membrane;

a diaphragm being positioned between the semipermeable membrane and the closed end;

a polymeric hydrogel enclosed between the semipermeable membrane and the diaphragm, the hydrogel including moieties that cause the hydrogel to change its swelling tendency and the pressure exerted by the hydrogel on the diaphragm in proportion to the free glucose concentration of the hydrogel;

an amount of a glucose-binding molecule immobilized in the hydrogel;

an amount of a hexose saccharide immobilized in the hydrogel;

a pressure transducer operatively engaged to the diaphragm; and

a battery powered telemeter operatively engaged to the transducer.

23. A biosensor for measuring the concentration of a free molecule, the biosensor comprising:

a polymeric hydrogel that changes its swelling tendency and pressure in proportion to the concentration of a free molecule;

a bound molecule immobilized in the hydrogel;

a high affinity and specific binding molecule chemically or physically immobilized in the hydrogel, configured such that the free molecule competitively binds with the bound molecule to the immobilized high affinity and specific binding molecule, thereby reducing the number of hydrogel crosslinks and causing the hydrogel to swell and increase the pressure of the hydrogel in the confined space;

a means for measuring the pressure or amount of swelling of the hydrogel; and

a means for reporting the concentration of the free molecule based on the measured pressure of the hydrogel.

24. A system for treating hypoglycemia, the system comprising:

a sensor for disposition in a patient, the sensor being configured to monitor blood glucose levels within a patient by use of a hydrogel;

alarm circuitry disposed in communication with the sensor for warning when the blood glucose level passes outside a predetermined parameter; and

an auto-injection device responsive to alarm signals for automatically injecting agents into the patient to return the blood glucose level to within the predetermined parameter.

25. A system for treating hypoglycemia, the system comprising:

a sensor for disposition in a patient, the sensor being configured to monitor blood glucose levels within a patient;

alarm circuitry disposed in communication with the sensor for warning when the blood glucose level passes outside a predetermined parameter;

an auto-injection device responsive to alarm signals for automatically injecting agents into the patient to return the blood glucose level to within the predetermined parameter; and

a global positioning system for determining the location of the patient.