# US 12,004,858 B2 · Analyte sensors employing multiple enzymes and methods associated therewith > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,004,858 B2 - Document ID: US12004858B2 - Status: Granted - Kind: B2 - Application number: 17/151,274 - Filed: 2021-01-18 - Published: 2021-05-13 - Granted: 2024-06-11 - Earliest claimed: 2019-01-28 - Assignees: Abbott Diabetes Care Inc. - Applicants: Abbott Diabetes Care Inc. - Inventors: Stephen Oja; Tianmei Ouyang; Hyun Cho; Lam N. Tran; Benjamin J. Feldman; Ashwin Kumar; Namvar Kiaie - Web page: https://patentgrep.com/patent/US12004858B2 ## CPC - A61B 5/14503: {invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors (A61B5/1459, A61B5/1464, A61B5/1473, A61B5/1482, A61B5/14865 take precedence)} - A61B 5/14865: {invasive, e.g. introduced into the body by a catheter or needle or using implanted sensors} - A61B 5/14532: {for measuring glucose, e.g. by tissue impedance measurement} - A61B 5/14546: {for measuring analytes not otherwise provided for, e.g. ions, cytochromes} - C12Q 1/002: {Electrode membranes} - G01N 27/3271: {Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood (amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001)} - C12Q 1/005: {involving specific analytes or enzymes (including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence)} - G01N 2333/9029: acting on -CH2- groups (1.17) - G01N 2333/904: acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1) - G01N 2333/908: acting on hydrogen peroxide as acceptor (1.11) - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer. ## Claims ### Claim 1 A method comprising: performing a dip coating operation to deposit a bilayer membrane upon a first active area disposed on a first working electrode of an analyte sensor, the first active area comprising an enzyme responsive to a first analyte, the bilayer membrane comprising an inner layer comprising a first membrane polymer and an outer layer comprising a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another; wherein the dip coating operation comprises one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer; depositing a homogeneous membrane upon a second active area disposed on a second working electrode of the analyte sensor, the second active area comprising an enzyme responsive to a second analyte, the homogeneous membrane comprising one of the first membrane polymer or the second membrane polymer; and wherein the inner layer of the bilayer membrane comprises a crosslinked polyvinylpyridine homopolymer or copolymer. ### Claim 2 (depends on 1) The method of claim 1, wherein the first active area is responsive to glucose or lactate. ### Claim 3 (depends on 1) The method of claim 1, wherein the first active area is responsive to lactate and the second active area is responsive to glucose. ### Claim 4 (depends on 3) The method of claim 3, wherein the bilayer membrane and the homogeneous membrane each comprise a crosslinked polyvinylpyridine homopolymer or copolymer. ### Claim 5 (depends on 1) The method of claim 1, wherein the homogeneous membrane and the outer layer of the bilayer membrane comprise the same membrane polymer. ### Claim 6 (depends on 1) The method of claim 1, wherein the homogeneous membrane and the inner layer of the bilayer membrane comprise the same membrane polymer. ### Claim 7 (depends on 1) The method of claim 1, wherein the first active area is responsive to ketones and the second active area is responsive to glucose. ### Claim 8 A method comprising: performing a dip coating operation to deposit (i) a bilayer membrane upon a first active area disposed on a first working electrode of an analyte sensor, the first active area comprising an enzyme responsive to a first analyte, and (ii) a homogeneous membrane upon a second active area disposed on a second working electrode of the analyte sensor, the second active area comprising an enzyme responsive to a second analyte; wherein the first active area and the second active area are laterally spaced apart from one another; wherein the first active area and the second active area are responsive to different analytes: wherein the bilayer membrane comprises an inner layer comprising a first membrane polymer and an outer layer comprising a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another; wherein the homogeneous membrane comprises one of the first membrane polymer or the second membrane polymer; and wherein the inner layer of the bilayer membrane comprises a crosslinked polyvinylpyridine homopolymer or copolymer. ### Claim 9 (depends on 8) The method of claim 8, wherein the homogeneous membrane and the outer layer of the bilayer membrane comprise the same membrane polymer. ### Claim 10 (depends on 8) The method of claim 8, wherein the homogeneous membrane and the inner layer of the bilayer membrane comprise the same membrane polymer. ### Claim 11 (depends on 8) The method of claim 8, wherein the first membrane polymer is deposited to define the inner layer, and the dip coating operation deposits the second membrane polymer upon the inner layer to define the outer layer and the homogeneous membrane. ### Claim 12 (depends on 8) The method of claim 8, wherein the first active area is responsive to lactate and the second active area is responsive to glucose. ### Claim 13 (depends on 12) The method of claim 12, wherein the bilayer membrane and the homogeneous membrane each comprise a crosslinked polyvinylpyridine homopolymer or copolymer. ### Claim 14 (depends on 8) The method of claim 8, wherein the first active area is responsive to ketones and the second active area is responsive to glucose. ### Claim 15 An analyte sensor comprising: at least a first working electrode and a second working electrode, a first active area disposed on the first working electrode and a second active area disposed on the second working electrode; wherein the first active area comprises an enzyme responsive to a first analyte and the second active area comprises an enzyme responsive to a second analyte; and wherein the first active area and the second active area are laterally spaced apart from one another; and a dip-coated bilayer membrane upon the first active area disposed on the first working electrode, and a dip-coated homogeneous membrane upon the second active area disposed on the second working electrode; wherein the bilayer membrane comprises an inner layer comprising a first membrane polymer and an outer layer comprising a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another; wherein the homogeneous membrane comprises one of the first membrane polymer or the second membrane polymer; and wherein the inner layer of the bilayer membrane comprises a crosslinked polyvinylpyridine homopolymer or copolymer. ### Claim 16 (depends on 15) The analyte sensor of claim 15, wherein the first active area is responsive to lactate and the second active area is responsive to glucose. ### Claim 17 (depends on 15) The analyte sensor of claim 15, wherein the analyte sensor is in communication with a control module and an electrical system of a vehicle, the control module including a computer system programmed to receive and process data provided by the analyte sensor, wherein operation of the vehicle is controlled or disabled by the computer system when a real-time measured analyte level of an operator crosses a predetermined safe threshold. ### Claim 18 (depends on 15) The analyte sensor of claim 15, wherein the first active area is responsive to ketones and the second active area is responsive to glucose.