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(12) United States Patent (10) Patent No.: US 7,163,616 B2 Vreelke Et Al

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(12) United States Patent (10) Patent No.: US 7,163,616 B2 Vreelke Et Al USOO71636.16B2 (12) United States Patent (10) Patent No.: US 7,163,616 B2 Vreelke et al. (45) Date of Patent: Jan. 16, 2007 (54) REAGENTS AND METHODS FOR (56) References Cited DETECTING ANALYTES, AND DEVICES U.S. PATENT DOCUMENTS COMPRISING REAGENTS FOR DETECTING ANALYTES 4,545,382 A 10/1985 Higgins et al. ............. 128,635 4,711,245 A 12/1987 Higgins et al. ... ... 128,635 4,863,016 A 9/1989 Fong et al. ........... ... 206,210 4.941,308 A 7, 1990 Grabenkort et al. .......... 53.425 (75) Inventors: Mark S. Vreeke, Houston, TX (US); 5, 120,420 A 6/1992 Nankai et al. .............. 204,403 Mary Ellen Warchal-Windham, 5,206,147 A 4, 1993 Hoenes ............. ... 435/25 Osceola, IN (US); Christina Blaschke, 5,212,092 A 5/1993 Jackson et al. ............... 436/11 White Pigeon, MI (US); Barbara J. 5,236,567 A 8/1993 Nanba et al. ............... 204,403 Mikel, Mishawaka, IN (US); Howard (Continued) A. Cooper, Elkhart, IN (US) FOREIGN PATENT DOCUMENTS (73) Assignee: Bayer Corporation, Elkhart, IN (US) EP O 330 517 A 8, 1989 (Continued) (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 OTHER PUBLICATIONS U.S.C. 154(b) by 102 days. Taylor, C.; Kenausis, G.; Katakis, I.; Heller, A.; “Wiring” of glucose oxidase within a hydrogel made with polyvinyl imidazole complexed with Ox-4,4'-dimethoxy-2,2'-bipyridin)CI+2+, Jour (21) Appl. No.: 10/231,539 nal of Electroanalytical Chemistry, 1995, vol. 396, pp. 511-515.* (Continued) (22) Filed: Sep. 3, 2002 Primary Examiner Kaj K. Olsen (74) Attorney, Agent, or Firm Jenkens & Gilchrist (65) Prior Publication Data US 2003 FOO94384 A1 May 22, 2003 (57) ABSTRACT Reagents for detecting an analyte are described. A reagent Related U.S. Application Data comprises (a) an enzyme selected from the group consisting (60) Provisional application No. 60/318,716, filed on Sep. of a flavoprotein, a quinoprotein, and a combination thereof. 14, 2001. and (b) a mediator selected from the group consisting of a phenothiazine, a phenoxazine, and a combination thereof. In (51) Int. C. addition, reagents having good stability to radiation steril GOIN 27/327 (2006.01) ization are described. Electrochemical sensors and sampling (52) U.S. Cl. ............................... 205/777.5; 204/403.14 devices comprising Such reagents, methods of producing a (58) Field of Classification Search ................ 204/403, sterilized device including Such reagents, and methods for 204/409, 412,403.14; 205/775, 777.5; 435/14, detecting an analyte which utilize Such reagents are 435/25, 26,810, 817: 436/806, 808, 149, described as well. 436/151 See application file for complete search history. 40 Claims, 6 Drawing Sheets 12 Layer 1 Layer 2 Layer3 Laminated Structure US 7,163,616 B2 Page 2 U.S. PATENT DOCUMENTS calcoaceticus'; Ph.D. Dissertation, Delft University of Technology, The Netherlands; 1999: pp. i-vi. 1-69. 5,298,144. A 3/1994 Spokane ..................... 204,403 5,334.508 A 8, 1994 Hoenes .... ... 435/25 Dewanti, Asteriani Ratih; "Enzymology of Quinoprotein Glucose 5,393,615 A 2, 1995 Corey et al. .................. 429.43 Dehydrogenases from Acinetobacter calcoaceticus'; Ph.D. Disser 5,411,647 A 5/1995 Johnson et al. .......... 205/777.5 tation, Delft University of Technology, The Netherlands; 2000; pp. 5,520,786 A 5/1996 Bloczynski et al. ... 204/403.14 i-viii. 1-107. 5,798,031 A 8, 1998 Charlton et al. ............ 204,403 Abstract of JP6114041A2 (issued Apr. 26, 1994): Jackson, Jeffrey 5,801,057 A 9, 1998 Smart et al. ...... ... 436/68 6,057,120 A 5, 2000 Heindl et al. .. ... 435/25 T.; Hui, Henry K. “Storage and Calibration Solution for Blood Gas 6,299,757 B1 * 10/2001 Feldman et al. ............ 205/775 Sensor Device of Blood in Multi-Charateristic Value Blood Vessel': 6,773,564 B1* 8/2004 Yugawa et al. ........ 204,403.14 pp. 1-2. Abstract of JP3180750A2 (issued Aug. 6, 1991): Yamaguchi, FOREIGN PATENT DOCUMENTS Hideichiro. "Cell for Chemical Sensor and Chemical Sensor Having EP 1 130 111 A 9, 2001 This Cell'; pp. 1-2. WO WO92 O7263 A 4f1992 Abstract of JP7083870A2 (issued Mar. 31, 1995): Kojima, Naomi; WO WO92 O7953. A 5, 1992 Sugama, Akio; Suzuki, Hiroaki. “Sterilization Method for Enzyme OTHER PUBLICATIONS Film”: pp. 1-2. Abstracts from ISI Current Contents, 1992-1999; 306 records. Olsthoorn, Arjen. "Structural and Mechanistic Aspects of Soluble Quinoprotein Glucose Dehydrogenase from Acinetobacter * cited by examiner U.S. Patent Jan. 16, 2007 Sheet 1 of 6 US 7,163,616 B2 12 Layer 1 Layer 2 Layer 3 Laminated Structure U.S. Patent Jan. 16, 2007 Sheet 2 of 6 US 7,163,616 B2 FG. 2 U.S. Patent Jan. 16, 2007 Sheet 3 of 6 US 7,163,616 B2 FIG. 3 Background Current After Irradiation 1400 1200 --Fas 1000 N S 800600 N a Control d 400 N :25kGy 200 .. N o50kGy O : N N 100 kGy NAD-MLB PQQ-Ferri PQQ-MLB Chemistry 600mg/dL Glucose Response After irradiation 18000 16000 S 1200014000 Že s SS %: a Control ° 33. NES 2666%.61%. f N 333. N D50is 100kéy kGy NAD-MLB POQ-Ferri PQQ-MLB Chemistry PQQ-GDH/MLB Dose Response Curve 20000 15000 g -0- 5 SeC. 5 10000 -H 10 SeC. as 5000 -A- 15 SeC. O O - 20 Sec. O 200 400 600 mg/dL Aqueous Glucose U.S. Patent Jan. 16, 2007 Sheet 4 of 6 US 7,163,616 B2 MLB 92 with Glucose Oxidase 3 25 a 20 S 15 CN y = 0.0423x + 0.53 (e) 105 R42 = 0.9934 9 S 0 C 0 100 200 300 400 500 600 glucose (mg/dL) GDH-PQQ With MLB 92 20000 0 initial 5 15000 4 wks at 50C E 10000 A RT 4Wk 40%RH 5 5000 no des O Linear (RT 4wk) O 200 400 600 Glucose (mg/dL) PQQ-GDHChemistry irradiation Stability Study Formulation 9. S 10000: -0- Control (S) 8 8000 --25kGy Sis 40006999 -A-50 kG y 3 2000 -o-100kGy g O O 200 400 600 mg/dL Aqueous Glucose U.S. Patent Jan. 16, 2007 Sheet 5 of 6 US 7,163,616 B2 PQQ-GDHChemistry irradiation Stability Study Formulation 20000 9 16000 (e) E. 12000 8000 -o-Control d 4000 --25kGy g O 0 1 00 200 300 400 500 600 mg/dL Aqueous Glucose PQQ-GDHChemistry irradiation Stability Study Os Formulation ill 3 3 20000 S 16000 9 oris 120008000 -o-Control d 4000 --25kGy KC O 0 1 00 200 300 400 500 600 mg/dL Aqueous Glucose PQQ-GDHChemistry irradiation Stability Study 2 Formulation IV 3 3 20000 S 16000 9 is 12000 5is de 80004000 --Control CDg O --25kGy 0 1 00 200 300 400 500 600 mg/dL Aqueous Glucose U.S. Patent Jan. 16, 2007 Sheet 6 of 6 US 7,163,616 B2 FIG. 12 PQQ-GDHChemistry irradiation Stability Formulation W O 93 20000 S 16000 s 12000 s: -o-Control O O --25kGy 0 1 00 200 300 400 500 600 mg/dL Aqueous Glucose US 7,163,616 B2 1. 2 REAGENTS AND METHODS FOR the radiation conditions commonly employed in lancet ster DETECTING ANALYTES, AND DEVICES ilization. Reagents stable to Such radiation sterilization COMPRISING REAGENTS FOR DETECTING could be incorporated into highly user-convenient units in ANALYTES which lancet and biosensor are combined. The present invention is directed to electron transfer This application claims benefit to provisional application mediators for use in flavoprotein- and quinoprotein-based 60/318,716, filed on Sep. 14, 2001. biosensor reagents, which exhibit improved stability to both environmental interferents and to radiation sterilization. BACKGROUND 10 SUMMARY The present invention relates to reagents, methods and devices for measurement of analytes and, more particularly, The scope of the present invention is defined solely by the to reagents, methods and devices for the measurement of appended claims, and is not affected to any degree by the glucose in the blood. statements within this Summary. By way of introduction, the The monitoring of certain analyte concentrations in the 15 presently preferred embodiments described herein are body enables early detection of health risks, and identifies directed towards remedying the aforementioned stability the need for the introduction of therapeutic measures. One of problems of electron transfer mediators and enzyme biosen the most commonly monitored analytes is glucose, the blood SOS. concentration of which is important in determining the Briefly stated, a composition aspect of the present inven appropriate dosages of insulin for diabetics. Various meth tion is directed to a reagent for detecting an analyte, com ods have been developed for monitoring glucose levels in prising (a) an enzyme selected from the group consisting of the blood, including the use of electrochemical biosensors. a flavoprotein, a quinoprotein, and a combination thereof. Electrochemical biosensors are based on enzyme-catalyzed and (b) a mediator selected from the group consisting of a chemical reactions involving the analyte of interest. In the phenothiazine, a phenoxazine, and a combination thereof. case of glucose monitoring, the relevant chemical reaction is 25 A first apparatus aspect of the present invention is directed the oxidation of glucose to gluconolactone. This oxidation is to an electrochemical sensor comprising: (a) a working catalyzed by a variety of enzymes, some of which may electrode having a Surface; and (b) a second electrode contain a bound coenzyme Such as nicotinamide adenine coupled to the working electrode. The surface of the work dinucleotide (phosphate) (NAD(P)), while others may con ing electrode is coated with a solution or mixture of a tain a bound cofactor Such as flavin adenine dinucleotide 30 reagent comprising an enzyme selected from the group (FAD) or pyrroloquinolinequinone (PQQ).
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