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Last Updated: March 28, 2024

Claims for Patent: 5,001,048


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Summary for Patent: 5,001,048
Title: Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film
Abstract:An electrical biosensor for analyte determination is prepared by polymerization of a mixture of a biological receptor capable of binding an analyte in a sample, a protein and a polymerizing agent such as glutaraldehyde to form a polymeric film on a transducer. The mixture preferably contains a stabilizer selected from lipids, detergents and antioxidents. The receptor may be an acetylcholine receptor and the analyte, acetylcholine. A preferred stabilizer is a combination of phosphatidyl choline and octyphenoxypolyethoxyethanol. In carrying out a determination, analyte in a sample binds to the receptor causing a change in an electrical characteristic of the film which is indicative of the presence of the analyte. The biosensor may contain a second polymeric film that is free of the receptor and which serves as a control.
Inventor(s): Taylor; Richard F. (Boxford), Marenchic; Ingrid G. (Walpole), Cook; Edward J. (South Hamilton, MA)
Assignee: Aurthur D. Little, Inc. (Cambridge, MA)
Application Number:07/058,389
Patent Claims:1. A method of immobilizing and stabilizing a biological receptor in a polymeric film on a transducer for the determination of an analyte in a sample, comprising:

(a) forming a mixture by combining a biological receptor capable of binding the analyte, a base component which is a protein, a polymerizing agent and at least one stabilizer selected from the group consisting of lipids, detergents and antioxidants, under conditions appropriate for polymerization to occur; and

(b) forming a film by polymerization of the mixture of (a) on a transducer.

2. A method of claim 1 wherein the protein is selected from the group consisting of bovine serum albumin, human serum albumin, gelatin and collagen; the polymerizing agent is glutaraldehyde; the lipid is selected from the group consisting of phosphatidylcholine, and cholesteryl palmitate, the detergent is selected from the group consisting of octylphenoxy polyethoxyethanol, sodium cholate, cetyltrimethylammonium bromide, polyoxyethylene sorbitan monooleate, N-decyl-N-dimethyl-3ammonio-1-propane sulfonate and N-octyl-N-dimethyl-3-ammonio-1-propane sulfonate and the antioxidant is selected from the group consisting of alpha-tocopherol and butylated hydroxyanisole.

3. A method for determining an analyte in a sample comprising:

(a) contacting the sample with a biosensor comprising:

(1) a polymeric film formed by chemical copolymerization of a solution comprising:

(a) a base component which is a protein

(b) a biological receptor capable of binding the analyte; and

(c) at least one stabilizer selected from the group consisting of lipids, detergents and antioxidants; and

(2) a sensor comprising a first conductor and a second conductor which are in contact with the polymeric protein film; and

(b) determining a change in an electrical characteristic of the polymeric protein film, whereby a change is indicative of the presence of analyte in the sample.

4. A biological receptor-containing biosensor for the determination of an analyte in a sample, comprising:

(a) a polymeric film formed by chemical copolymerization of a solution comprising

(1) a base component which is a protein;

(2) a biological receptor capable of binding the analyte; and

(3) at least one stabilizer selected from the group consisting of lipids, detergents and antioxidants;

(b) a first sensor comprising a first conductor and a second conductor which are in contact with said film; and

(c) a current source in conductive contact with the first and second conductors for measuring an electrical characteristic of the polymeric film.

5. A biological receptor-containing biosensor of claim 4 wherein the biological receptor is a neurological receptor.

6. A biological receptor-containing biosensor of claim 5 wherein the neurological receptor is an acetylcholine receptor.

7. A biological receptor-containing biosensor of claim 4 wherein the protein is selected from the group consisting of serum albumin, gelatin and collagen.

8. A biological receptor-containing biosensor of claim 4 wherein the lipid is selected from the group consisting of: phosphatidyl choline and cholesteryl palmitate, the detergent is selected from the group consisting of octylphenoxy polyethoxethanol, sodium cholate, cetyltrimethyl-ammonium bromide, polyoxyethylene, sorbitan monooleate, N-decyl-N-dimethyl-3-ammonio-1-propane sulfonate and N-octyl-N-dimethyl-3-ammonio-1- propane sulfoante, and the antioxidant is selected from the group consisting of alpha-tocopherol and butylated hydroxyanisole.

9. An acetylcholine receptor-containing biosensor for the determination of an analyte in a sample comprising:

(a) a polymeric film formed by chemical copolymerization of a solution comprising

(1) a base component which is a protein selected from the group consisting of serum albumin, gelatin and collagen;

(2) acetycholine receptor capable of binding the analyte; and

(3) at least one stabilizer selected from the group consisting of lipids, detergents and antioxidants;

(b) a first sensor comprising a first conductor and a second conductor which are in contact with said film; and

(c) a current source in conductive contact with the first and the second conductors for measuring an impedance of the film.

10. A receptor of claim 9 wherein the stabilizer is a combination of the lipid, phosphatidyl choline, and the detergent, octylphenoxypolyethoxyethanol.

11. A biological receptor-containing biosensor for the determination of an analyte in a sample, comprising a first polymeric film comprising a protein, a stabilizer selected from the group consisting of lipids, detergents and antioxidants and a biological receptor capable of binding the analyte, the first polymeric film being in contact with a first electrical sensor for measuring an electrical characteristics of the first polymeric film and further comprising:

(a) a second polymeric film free of receptor for binding the analyte, the second polymeric film being in contact with a second electrical sensor for measuring an electrical characteristic of the second polymeric film;

(b) a potential barrier interposed between the second polymeric film and the first polymeric film, such that the barrier hinders current flow between said second polymeric film and the first polymeric film;

(c) a current source; and

(d) a circuit contacting the current source and the first and the second electrical sensors for measuring absolute and the differential changes in the electrical characteristics of the first and second films.

12. A biological receptor-containing biosensor of claim 11 wherein the biological receptor is a neutrological receptor.

13. A biological receptor-containing biosensor of claim 12 wherein the neurological receptor is an acetylcholine receptor.

14. A biological receptor-containing biosensor of claim 11 wherein the protein is selected from the group consisting of serum albumin, gelatin and collage.

15. A biological receptor-containing biosensor of claim 11 wherein the lipid is selected from the group consisting of: phosphatidyl choline and cholesteryl palmititate, the detergent is selected from the group consisting of octylphenoxypolyethoxethanol, sodium cholate, cetyltrimethyl-ammonium bromice, polyoxyethylene, sorbitan monooleate, N-decyl-N-dimethyl-3-ammonio-1-propane sulfonate and N-octyl-N-dimethyl-3-ammonio-1-propane sulfonate and the antioxidant is selected from the group consisting of alpha-tocopherol and butylated hydroxyanisole.

Details for Patent 5,001,048

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Grifols Therapeutics Llc ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5 albumin (human) For Injection 101138 10/21/1942 ⤷  Try a Trial 2040-01-28
Baxalta Us Inc. BUMINATE, FLEXBUMIN albumin (human) Injection 101452 03/03/1954 ⤷  Try a Trial 2040-01-28
Csl Behring Ag ALBURX albumin (human) Injection 102366 07/23/1976 ⤷  Try a Trial 2040-01-28
Grifols Biologicals Llc ALBUTEIN albumin (human) Injection 102478 08/15/1978 ⤷  Try a Trial 2040-01-28
Instituto Grifols, S.a. HUMAN ALBUMIN GRIFOLS albumin (human) Injection 103352 02/17/1995 ⤷  Try a Trial 2040-01-28
Instituto Grifols, S.a. HUMAN ALBUMIN GRIFOLS albumin (human) Injection 103352 06/11/2003 ⤷  Try a Trial 2040-01-28
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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