Hydrogel biosensor

• US 6,514,689 B2
• Filed: 04/20/2001
• Issued: 02/04/2003
• Est. Priority Date: 05/11/1999
• Status: Active Grant

• Current Assignee: M-Biotech, Inc.
• Original Assignee: M-Biotech, Inc.
• Inventors: Han, In Suk, Lew, Seok Lew, Magda, Jules John, Jean, Young San
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/839,993
• Publication Number:
  US 20020042065A1
• Time in Patent Office: 655 Days
• Field of Search: 435/4, 435/14, 435/25, 435/287.1, 435/287.7, 436/95, 436/148, 604/891.1, 604/892.1
• US Class Current: 435/4
• CPC Class Codes: A61B 5/0022 Monitoring a patient using a global network, e.g. telephone networks, internet
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/14865 invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors
• 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 2001/021 Correlating sampling sites with geographical information, e.g. GPS
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 7/00 Analysing materials by measuring the pressure or volume of a gas or vapour

Abstract

A biosensor (10) has a hydrogel (30) in a rigid and preferably biocompatible enclosure (20). The hydrogel (30) includes an immobilized analyte binding molecule (ABM) and an immobilized analyte. The immobilized analyte competitively binds with free analyte to the ABM, thus changing the number of crosslinks in the hydrogel (30), which changes hydrogel swelling tendency (and thus the osmotic pressure) in its confined space in proportion to the concentration of free analyte concentration. By measuring the change in hydrogel pressure with a pressure transducer (40), the biosensor (10) is able to accurately measure the concentration of the free analyte 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 analyte level is outside desired predetermined parameters, and/or to automatically inject an agent to counteract the adverse analyte levels.

Claims

1. A biosensor for measuring the concentration of molecules of an analyte in a body fluid, comprising:

a polymeric hydrogel having pendant moieties that are charged under physiological conditions;

an analyte binding molecule immobilized in the hydrogel and capable of binding the free analyte;

analyte molecules immobilized in the hydrogel; and

pressure detection means for measuring the osmotic pressure of the hydrogel.

2. The biosensor of claim 1, wherein the hydrogel is disposed within a rigid enclosure with at least one area permeable to contact between the hydrogel and a fluid being tested and permitting free analyte molecules to diffuse into the hydrogel from the fluid.

3. The biosensor of claim 1, wherein the pressure detection means comprises a diaphragm disposed within the enclosure in contact with the hydrogel, and a pressure transducer operably engaged with the diaphragm to measure pressure on the diaphragm.

4. The biosensor of claim 1, further including reporting means operably associated with the pressure-detection means for reporting a data signal reflective of the pressure changes in the hydrogel.

5. The biosensor of claim 2, wherein the permeable area is an open end in the rigid enclosure, and said open end is sealed by a semipermeable membrane that allows the free analyte molecules to diffuse into the hydrogel.

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

7. The biosensor of claim 4 wherein the enclosure is coated with a semipermeable membrane and a biodegradable polymer on the semipermeable membrane.

8. The biosensor of claim 1 wherein the analyte binding molecule is selected from the group consisting of:

antibodies, enzymes, membrane receptors, kinases, Protein A, Poly U, Poly A, Poly lysine, triazine dye, nucleoside, boronic acid, thiol, heparin, polysaccharides, Coomassie blue, azure A, and metal-binding peptides, proteins, and chelating agents.

9. The biosensor of claim 1 wherein the immobilized analyte is selected from the group consisting of:

antigens, enzyme cofactors, enzyme substrates, enzyme inhibitors, IGG, sugar, carbohydrate, nucleic acids, nucleotide, nucleoside, cysteine, arginine, lysine, protamine, heparin, dyes, and metal ions.

10. The biosensor of claim 1, wherein the charged pendant groups are present at a density chosen to optimize the amount of hydrogel swelling in response to changes in level of free analyte molecules.

11. The biosensor of claim 1, wherein the immobilized analyte molecules and immobilized analyte binding molecule are present at respective densities chosen to optimize the amount of hydrogel swelling in response to changes in level of free analyte molecules.

12. The biosensor of claim 3 wherein the pressure transducer is selected from the group consisting of a piezo-electric transducer, a piezoresistive transducer, and a capacitive transducer.

13. The biosensor of claim 4, further including computing means connected to receive the data signal from the reporting means, said computing means being constructed to compare the data signal to a calibration curve to compute a concentration of the free analyte in the body fluids and produce an output signal representing the free analyte concentration.

14. The biosensor of claim 4, wherein the reporting means is a battery powered telemeter, and further including receiving means positioned at a location remote to the patient for receiving the data signal.

15. The biosensor of claim 14, further including computing means operably associated with the receiving means for comparing the data signal to a calibration curve to compute a concentration of the free analyte in the body fluids and produce an output signal representing the analyte concentration.

16. The biosensor of claim 14, wherein the computer means is further configured to compare the detected analyte concentration to a predetermined safe range, and to produce an alarm signal when the detected analyte concentration falls outside the safe range.

23. The biosensor of claim 1, wherein the pressure detection means for measuring the osmotic pressure of the hydrogel reports a data signal reflective of the pressure of the hydrogel, and additionally including means for generating an alarm signal responsive to the reported data signal if outside a predetermined range, and a transmitter for transmitting the alarm signal to a remote location.

24. The biosensor of claim 23, 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.

25. The biosensor of claim 23, additionally including an injection device for injecting agents into a person in response to alarm signals from the means for generating an alarm signal.

26. The biosensor of claim 23, additionally including an injection device for injecting agents into a person in response to reported data signal.

27. The biosensor of claim 23, additionally including a GPS unit operably associated with the transmitter for transmitting a location signal reflective of a patient'"'"'s location in addition to the alarm signal.

17. A method of determining the concentration of free analyte in a solution, comprising the steps of:

providing a hydrogel having pendant charged moieties, analyte molecules, and analyte-specific binding molecules covalently immobilized therein;

enclosing the hydrogel in a rigid structure which has at least one permeable portion available for contacting a test fluid with the hydrogel, the permeable portion constructed to permit free analyte to diffuse into the hydrogel;

contacting the hydrogel sequentially with a series of calibration solutions having known concentrations of free analyte;

measuring osmotic pressure in the hydrogel for each of the calibration solutions to produce a calibration curve of osmotic pressure versus analyte concentration;

contacting the hydrogel with the test fluid, and measuring a resulting osmotic pressure; and

comparing the resulting osmotic pressure with the calibration curve to determine analyte concentration of the test fluid.

18. The method of claim 17, wherein said steps of measuring the osmotic pressure are accomplished by disposing pressure sensing means within the rigid structure and in contact with the hydrogel for measuring osmotic pressure of the hydrogel and producing a data signal reflective thereof.

19. The method of claim 18, wherein said pressure sensing means comprises a diaphragm disposed within the rigid enclosure in contact with the hydrogel, and a pressure transducer operably engaged with the diaphragm to measure pressure on the diaphragm.

20. A sensor for measuring the concentration of free molecules of an analyte in a fluid, comprising:

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

a diaphragm positioned within the enclosure between the semipermeable membrane and the closed end;

a polymeric hydrogel having pendant moieties which are charged at physiological pH, the hydrogel being enclosed between the semipermeable membrane and the diaphragm such that changes in osmotic pressure within the hydrogel are accompanied by changes in pressure exerted on the diaphragm;

analyte binding molecules immobilized within the hydrogel;

analyte molecules immobilized within the hydrogel; and

a pressure transducer operatively engaged to the diaphragm.

21. The sensor of claim 20, further including a battery powered telemeter operatively engaged to the transducer.

22. A method for using a biosensor to measure the concentration of free molecules of an analyte in a fluid, including:

a first step of providing a 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 positioned between the semipermeable membrane and the closed end such that changes in osmotic pressure within the hydrogel are accompanied by changes in pressure exerted on the diaphragm;

a polymeric hydrogel having pendant moieties which are charged at physiological pH, the hydrogel being enclosed between the semipermeable membrane and the diaphragm such that changes in osmotic pressure within the hydrogel are accompanied by changes in pressure exerted on the diaphragm;

analyte binding molecules immobilized within the hydrogel;

analyte immobilized within the hydrogel, and

osmotic pressure sensing means operatively engaged to the diaphragm for sensing the osmotic pressure exerted thereon and providing a data signal reflective thereof;

a second step of providing computing means connected to receive the data signal, compare it to a predetermined calibration curve of osmotic pressure vs, concentration of free analyte molecules, and output a concentration value;

a third step of inserting the biosensor into the fluid and allowing sufficient time for free analyte molecules to diffuse to equilibrium within the hydrogel; and

a fourth step of reading the concentration value output by the computing means.