EP2959008A1 - Fabrication à haut rendement de bandelettes de test du glucose sanguin - Google Patents

Fabrication à haut rendement de bandelettes de test du glucose sanguin

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Publication number
EP2959008A1
EP2959008A1 EP14705543.8A EP14705543A EP2959008A1 EP 2959008 A1 EP2959008 A1 EP 2959008A1 EP 14705543 A EP14705543 A EP 14705543A EP 2959008 A1 EP2959008 A1 EP 2959008A1
Authority
EP
European Patent Office
Prior art keywords
batch
diagnostic
detection reagent
coenzyme
chemical detection
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP14705543.8A
Other languages
German (de)
English (en)
Inventor
Claudia Gaessler-Dietsche
Hans-Peter Haar
Carina HORN
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
F Hoffmann La Roche AG
Roche Diabetes Care GmbH
Original Assignee
F Hoffmann La Roche AG
Roche Diagnostics GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by F Hoffmann La Roche AG, Roche Diagnostics GmbH filed Critical F Hoffmann La Roche AG
Priority to EP14705543.8A priority Critical patent/EP2959008A1/fr
Publication of EP2959008A1 publication Critical patent/EP2959008A1/fr
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/54—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving glucose or galactose
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/902—Oxidoreductases (1.)
    • G01N2333/904—Oxidoreductases (1.) acting on CHOH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)

Definitions

  • the present invention relates to a method for the production of diagnostic elements, a diagnostic product comprising a stable chemical detection reagent, and the use of this stable chemical detection reagent.
  • Diagnostic elements are important components of clinically relevant analysis methods.
  • analytes e.g. Metabolites or substrates in the foreground, which are determined directly or indirectly, for example, using an enzyme specific for the analyte.
  • the analytes are here reacted with the aid of an enzyme-coenzyme complex and then quantified using suitable means.
  • the analyte to be determined is brought into contact with a suitable enzyme and a coenzyme, the enzyme usually being used in catalytic amounts.
  • the coenzyme is physicochemically altered by the enzymatic reaction, e.g. oxidized or reduced, and detects the process, for example, electrochemical or photometric.
  • a calibration provides a direct correlation between the measured value and the concentration of the analyte to be determined.
  • the error results are based primarily on the fact that the substances used in such diagnostic elements, in particular enzymes, coenzymes or / and mediators, generally sensitive to moisture, heat and / or light and are inactivated over time. This has u.a. As a result, during the manufacturing process certain service lives of a test chemical prepared in aqueous solution must not be exceeded and the test chemical applied to a suitable carrier can not be stored for an excessively long time before the next processing step.
  • test chemistry To accommodate these limitations dictated by test chemistry, the size of batches of a test chemistry used to make diagnostic elements is usually limited. In this way it can be ensured that a single batch of the test chemistry is substantially homogeneous and that the substances contained in the test chemistry furthermore have a sufficiently high activity for the subsequent detection of an analyte to be determined even after their processing into a diagnostic element.
  • the object underlying the invention was thus to a To provide methods for the production of diagnostic elements, in which the disadvantages of the prior art are at least partially eliminated.
  • the method should enable the provision of large and homogeneous batches of test chemistry to be used, ensure high homogeneity of sequentially produced batches of diagnostic elements, allow the use of a single batch coding for a large number of diagnostic elements, and, moreover, the possibility of encoding individual diagnostic elements Offer.
  • the second diagnostic elements are each provided with the batch coding produced in step (d) before the separation of the coated second carrier.
  • a chemical detection reagent comprising a combination of a coenzyme-dependent enzyme and an artificial coenzyme greatly simplifies the preparation, testing, packaging and handling of diagnostic elements due to its high stability to moisture, heat and light. Furthermore, the use of such a chemical detection reagent results in time and financial advantages, which are of paramount importance in the large-scale production of diagnostic test elements.
  • the first step of the method of the invention requires the provision of a batch of a chemical detection reagent which i.a. a coenzyme-dependent enzyme, such as a flavin, nicotinamide or pyrroloquinoline quinone-dependent oxidoreductase.
  • a coenzyme-dependent enzyme such as a flavin, nicotinamide or pyrroloquinoline quinone-dependent oxidoreductase.
  • the method according to the invention provides for the use of a flavin, nicotinamide or pyrroloquinoline quinone-dependent dehydrogenase, with nicotinamide-dependent dehydrogenases, in particular NAD (P) / NAD (P) H-dependent
  • a coenzyme-dependent enzyme is preferably a dehydrogenase selected from the group consisting of alcohol dehydrogenase (EC 1.1.1.1, EC 1.1.1.2), L-amino acid dehydrogenase (EC 1.4.1.5), glucose dehydrogenase (EC 1.1.1.47 ), Glucose-6-phosphate
  • Dehydrogenase (EC 1.1.1.49), glycerol dehydrogenase (EC1 .1.6), 3-hydroxybutyrate dehydrogenase (EC 1.1.1.30), lactate dehydrogenase (EC 1.1.1.27, EC 1.1.1.28), malate dehydrogenase ( EC 1.1.1.37), glutamate dehydrogenase (EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4) and sorbitol dehydrogenase. More preferred is the enzyme glucose dehydrogenase (E.C. 1.1.1.47), glucose-6-phosphate
  • mutated coenzyme-dependent enzyme designate a genetically modified variant of a native coenzyme-dependent enzyme (wild-type enzyme), which has the same number of amino acids the wild-type enzyme has altered amino acid sequence, that is different in at least one amino acid from the wild-type enzyme.
  • wild-type enzyme a native coenzyme-dependent enzyme
  • the mutant preferably has an increased thermal or / and hydrolytic stability compared with the wild-type enzyme.
  • the mutant coenzyme-dependent enzyme can be obtained by mutation of a native coenzyme-dependent enzyme derived from any biological source, the term "biological source” for the purposes of this invention encompassing both prokaryotes and eukaryotes site-specific or non-site-specific, preferably site-specific using recombinant methods known in the art, wherein at least one amino acid exchange results within the amino acid sequence of the native enzyme.
  • the coenzyme-dependent enzyme used is a mutated glucose dehydrogenase (EC 1.1.1.47) or a mutated glucose-6-phosphate dehydrogenase (EC 1.1.1.49).
  • mutated glucose dehydrogenases are described, inter alia, in WO 2005/045016, WO 2011/020856, Baik (Appl. Environ. Microbiol. (2005), 71, 3285) and Väsquez-Figueroa (ChemBioChem (2007), 8, 2295) , the disclosure of which is hereby incorporated by reference.
  • the coenzyme-dependent enzyme is a mutant glucose dehydrogenase having the amino acid sequence shown in SEQ ID NO: 1 (GlucDH_E96G_E170K) or SEQ ID NO: 2 (GlucDH_E170K_K252L).
  • the chemical detection reagent used according to the invention furthermore comprises an artificial coenzyme.
  • An artificial coenzyme for the purposes of the present invention is a coenzyme chemically modified with respect to the native coenzyme, which at atmospheric pressure has a higher stability to moisture compared to native coenzyme, temperatures in particular in the range from 0 ° C. to 50 ° C., acids and bases, in particular Range of pH 4 to pH 10, and / or nucleophiles such as alcohols or amines, and thus can operate under identical environmental conditions over a longer period than the native coenzyme its effect.
  • the artificial coenzyme has a higher hydrolytic stability compared to the native coenzyme, with full hydrolytic stability being particularly preferred under test conditions.
  • the artificial coenzyme may have a reduced binding constant for the coenzyme-dependent enzyme, for example a binding constant reduced by a factor of 2 or more.
  • artificial coenzymes which can be used in the context of the process according to the invention are artificial NAD (P) / NAD (P) H compounds, ie chemical derivatives of native nicotinamide adenine dinucleotide (NAD / NADH) or native nicotinamide Adenine dinucleotide phosphate (NADP / NADPH) or the compound of formula (I)
  • the NAD (P) / NAD (P) H artificial compound preferably comprises a 3-pyridinecarbonyl or a 3-pyridinethiocarbonyl -Rest, which is linked without glycosidic bond via a linear or cyclic organic radical, in particular via a cyclic organic radical, with a phosphorus-containing radical, such as a phosphate radical.
  • the artificial coenzyme is particularly preferably selected from a compound of the general formula (II):
  • A adenine or an analog thereof
  • T each independently O, S,
  • V each independently OH or a phosphate group, or two Groups that form a cyclic phosphate group;
  • X 1 , X 2 each independently O, CH 2 , CHCH 3 , C (CH 3 ) 2 , NH, NCH 3 ,
  • Y NH, S, O, CH 2 ,
  • Z a linear or cyclic organic radical
  • Z is particularly preferably a saturated or unsaturated carbocyclic or heterocyclic five-membered ring, in particular a radical of the general formula (III),
  • R 4 each independently H, F, Cl, CH 3 ,
  • R 6 , R 6 ' each independently CH or CCH 3 .
  • the compounds of general formula (II) contain an adenine analog.
  • adenine analog refers to a chemical derivative of native adenine that exhibits the same pharmacological activity in the human body as adenine.
  • "Concrete examples of adenine analogs include, in particular, C" 8 "and N" 6 " -substituted adenine, 7- deazaadenine, 8-azaadenine, 7-deaza-8-azaadenine and formycin, wherein the 7-Deazaversionn in the 7-position with halogen, Ci -6 alkynyl, Ci -8 alkenyl or Ci-e Alkyl may be substituted.
  • the compounds contain adenosine analogues which instead of ribose contain, for example, 2-methoxydesoxyribose, 2'-fluorodeoxyribose, hexitol, altritol or polycyclic analogs, such as bicyclo, LNA and tricyclo sugars.
  • ribose contain, for example, 2-methoxydesoxyribose, 2'-fluorodeoxyribose, hexitol, altritol or polycyclic analogs, such as bicyclo, LNA and tricyclo sugars.
  • ribose contain, for example, 2-methoxydesoxyribose, 2'-fluorodeoxyribose, hexitol, altritol or polycyclic analogs, such as bicyclo, LNA and tricyclo sugars.
  • di) -phosphate oxygens isotronically for example O " by S " or BH 3 " , O by
  • the artificial coenzyme is the compound known in the literature as carbaNAD (JT Slama, Biochemistry (1988), 27, 183 and Biochemistry (1989), 28, 7688).
  • the chemical detection reagent may comprise other substances which serve for the qualitative detection or / and the quantitative determination of analytes, such as a mediator and / or an optical indicator.
  • mediator refers to a chemical compound which increases the reactivity of the reduced coenzyme obtained by reaction with the analyte and allows for the transfer of electrons to a suitable optical indicator or optical indicator system
  • mediators include in particular azo compounds, nitrosoanilines, quinones and phenazines.
  • any substance which is reducible and undergoes a visually and / or machine-detectable change in its optical properties can be used as the optical indicator.
  • Preferred optical indicators for the purposes of the present invention include reducible heteropolyacids, in particular 2,18-phosphomolybdic acid.
  • quinones such as resazurin, dichlorophenolindophenol or / and tetrazolium salts, can be used as optical indicators.
  • a first part of this charge is applied to a first carrier, such as a film or an injection molded part, and then dried, thereby coating the first carrier with the first batch of chemical detection reagent , While both this step and subsequent processing steps using conventional test chemistry which is sensitive to moisture, heat and / or light may only take a short time, it can be achieved by the use of a combination of coenzyme-dependent enzyme and artificial coenzyme and thereby achieved long-term stability of the test chemistry can be dispensed with an immediate further processing of the complete batch of the chemical detection reagent, which brings significant production advantages.
  • a single batch of the chemical detection reagent can also coat a larger number of supports, ultimately allowing the generation of a larger batch of diagnostic elements which is also more homogeneous than a single batch of diagnostic elements comprising conventional test chemistry.
  • the chemical detection reagent described above has the advantage that multiple batches of diagnostic elements can be generated by means of a single batch, which are then homogeneous with each other. In this way, it is ensured that even with a large batch of the chemical detection reagent, the first diagnostic elements produced therefrom and the last are identical or merely vary within narrow limits. In contrast, when using conventional test chemistry, small batches of diagnostic test elements have to be produced so that the dispersion of the reactivity of the chemical detection reagent, which deteriorates due to environmental influences, still remains within the permissible range.
  • the coated first carrier is separated by means of suitable techniques into a plurality of first diagnostic elements (preliminary batch) and, using at least one of the first diagnostic elements, a batch coding is generated for the complete batch of the chemical detection reagent.
  • the batch coding produced in this context preferably contains a mathematical equation which indicates the optionally temperature-dependent relationship between the respective amount of an analyte to be determined and the resulting signal, which can be measured optically or electrochemically, for example.
  • any code which appears appropriate to the person skilled in the art for the coding of diagnostic elements and which allows reliable traceability of the diagnostic elements provided with the code can be used as batch coding.
  • an optically and / or electronically readable code is used as batch coding, such as a barcode or an RFID transponder.
  • a barcode which can be provided in one or two dimensions, can be configured in black-and-white, in grayscale or in color and, if desired, can comprise a hologram.
  • the method according to the invention provides for the coating of a second carrier, in particular a film or an injection-molded part, with a second part of the batch of the chemical detection reagent.
  • a coated second support is obtained, from which, by subsequent singulation, a plurality of second diagnostic elements are obtained, which can be subjected to review as necessary and suitably packaged after passing quality control.
  • the coding of the second diagnostic elements takes place according to the invention before the separation of the coated second carrier.
  • the second diagnostic elements are each provided individually with the batch coding based on the preliminary batch (ie with the first diagnostic elements), preferably by applying the batch coding to the second carrier before coating the second carrier with the second part of the batch chemical detection reagent is realized.
  • This has considerable financial and production advantages, since a single batch coding is sufficient to encode an amount of diagnostic elements increased by a factor of 2, preferably by a factor of 3, particularly preferably by a factor of 5, as in the case of conventional test chemistry.
  • the method according to the invention makes it possible to code each individual diagnostic element by targeted application of the batch coding to the second carrier, which is generally not possible when using conventional test chemistry.
  • the preparation of batch coding usually requires the preparation of a pre-batch.
  • the subsequent coding of diagnostic elements with the batch coding obtained in this case must be carried out in an earlier production step, and in in any case prior to separation of the respective carrier to the diagnostic elements, to be economically justifiable and technically feasible.
  • coding individual diagnostic elements By realizable according to the invention coding individual diagnostic elements, the use of ROM keys or the input of code numbers by the consumer is dispensable.
  • the advantage of encoding individual diagnostic elements over the encoding of an entire magazine e.g., a can of test strips
  • an inappropriate code e.g., with an earlier batch of the diagnostic element
  • a batch of the chemical detection reagent need not be further processed immediately after its preparation in order to avoid a change in its chemical and / or physical properties. Rather, the inventive method allows that between the first Providing a batch of the chemical detection reagent and its complete processing into diagnostic elements can be from several hours to several weeks, offering both temporal and production advantages.
  • a period of at least 36 hours, preferably at least 48 lies between the provision of the charge of the chemical detection reagent and the subsequent coating of the second carrier with the second part of the batch of chemical detection reagent hours.
  • a period of at least 20 days, preferably of at least 30 days lies between the coating of the second carrier with the second part of the batch of the chemical detection reagent and the separation of the coated second carrier into a plurality of second diagnostic elements ,
  • the period between the provision of the batch of the chemical detection reagent and the coating of the second support, or / and between the coating of the second support and its separation into a plurality of second diagnostic elements is preferably selected such that the batch of the chemical detection reagent is substantially free Changes in their chemical and / or physical properties and thus enables the production of diagnostic elements, which ensure a determination of analytes within the legally permissible limits.
  • substantially no change in chemical and / or physical properties in this context means a decrease in the activity of the coenzyme-dependent enzyme or a decrease in the content of artificial coenzyme in the chemical detection reagent of less than 40%, preferably less than 30%, and most preferably less than 20%, based on the activity of the coenzyme dependent enzyme and based on the content of artificial coenzyme in the chemical detection reagent immediately after its production.
  • the first diagnostic elements and second diagnostic elements generated by means of the method according to the invention can be identical or different, but are preferably of the same design.
  • the diagnostic elements may, in principle, have any physical form known to those skilled in the art, which is suitable for determining the presence and / or amount of an analyte in a sample, and each comprise at least one test field, which are brought into contact with a sample containing the analyte can and with the aid of suitable ittel a qualitative or / and quantitative determination of the analyte allows.
  • first diagnostic elements and / or second diagnostic elements include, in particular, test strips, test tapes and test discs from which diagnostic elements based thereon, such as test strip magazines and tape magazines, can be made, if necessary.
  • Suitable test strip magazines include in particular blister magazines, Leporellomagazine, disc magazines, staple magazines, drum magazines and turning magazines, which u.a. in EP 0 951 939, EP 1 022 565, EP 1 736 772, WO 2005/104948 and WO 2010/094427.
  • Tape magazines are u.a. from DE 10 2005 013 685, EP 1 739 432 and WO 2004/047642. The disclosure of the above publications is hereby incorporated by reference.
  • the first diagnostic elements and / or the second diagnostic elements each comprise a plurality of test fields, preferably at least 10 individual test fields, more preferably at least 25 individual test fields, and most preferably at least 50 individual ones Test fields.
  • the individual test fields are each arranged at a distance of a few millimeters to a few centimeters, for example at a distance of ⁇ 2.5 cm, from each other.
  • each of the test fields of the first diagnostic elements and / or the second diagnostic elements is at least partially surrounded by a hydrophobic edge or / and is stored in its own, substantially closed chamber.
  • a hydrophobic edge is particularly advantageous in cases in which a consumer has to apply a sample of the analyte to be determined manually to a test field of the respective diagnostic element and an overflow of the sample to adjacent test fields or an entry of the sample into a for measuring the diagnostic element required device is to be avoided.
  • Diagnostic elements in which each test field is stored in its own, essentially closed chamber are used, for example, in fully automatic measuring systems.
  • Such measuring systems have several test fields, e.g. can be arranged annularly side by side on a rotatable plate, as well as many trained to scrape skin needle elements in magazinierter form.
  • no manual application of the sample to a test field has to be made by the consumer (this is done automatically by the system), the individual test fields must be in separate compartments for reasons of hygiene.
  • substantially closed chamber in this context means that the chamber walls can be permeable to air and / or water, but do not allow the entry of dust particles into the chamber and thus ensure a dust-free storage of the test fields.
  • test fields of a diagnostic element conventional test chemistry, which has a low stability to moisture, heat and / or light, so must the test fields at least partially surrounding hydrophobic edge or the enclosing the test fields chambers must be prepared in a complicated and technically complex process.
  • the chemical detection reagent used according to the invention is stable to, inter alia, light, and in particular UV light
  • the diagnostic elements described in the present application may also comprise photocurable or UV-curable materials.
  • the method according to the invention provides in a preferred variant that the hydrophobic edge at least partially surrounding the test fields of a diagnostic element and / or the chamber walls of the chambers enclosing the individual test fields are formed from a UV-curable material.
  • UV-curable materials which may be used in this context are known to those skilled in the art and include i.a. Epoxy and acrylate adhesives, but are not limited to these.
  • the method according to the invention additionally comprises a check of the thickness and / or the homogeneity of the layer of the chemical detection reagent on the second diagnostic elements.
  • diagnostic elements which are coated with a chemical detection reagent used according to the invention also have a high stability to moisture. Accordingly, diagnostic elements which have been produced by means of the method according to the invention and subsequently packaged are preferably not subjected to a water vapor tightness test, whereby the entire production process can be accelerated and the production costs can be reduced.
  • the diagnostic elements produced by the method according to the invention can be used for the qualitative and / or quantitative determination of any biological or chemical substance which can be detected optically or electrochemically.
  • the analyte is selected from the group consisting of malic acid, alcohol, ascorbic acid, cholesterol, glucose, glycerol, urea, 3-hydroxybutyrate, lactic acid, pyruvate and triglycerides, with glucose being particularly preferred.
  • the analyte to be determined may be from any source, but is preferably in a body fluid, including, but not limited to, whole blood, plasma, serum, lymph, bile, cerebrospinal fluid, extracellular tissue fluid, urine, and glandular secretions such as saliva or sweat, contain.
  • a body fluid including, but not limited to, whole blood, plasma, serum, lymph, bile, cerebrospinal fluid, extracellular tissue fluid, urine, and glandular secretions such as saliva or sweat, contain.
  • the presence or / and the amount of an analyte in a sample selected from the group consisting of whole blood, plasma and serum is preferably determined by means of the detection reagent according to the invention.
  • the invention relates to a diagnostic product comprising (a) at least one diagnostic element, and (b) optionally at least one needle element for scoring skin.
  • the diagnostic element in turn comprises (i) a carrier, (ii) a chemical detection reagent comprising a coenzyme-dependent enzyme and an artificial co
  • the diagnostic product according to the invention in addition to the chemical detection reagent further comprises a needle element for scoring skin, which preferably consists of a sterilizable material, such as metal or plastic.
  • the needle element preferably comprises a capillary channel, by means of which a sufficient amount of the sample for the determination of the analyte and taken by taking capillary forces on a test field of the diagnostic element can be applied.
  • diagnostic element including the preferred variants for carrier, chemical detection reagent and batch coding
  • the invention features the use of a chemical detection reagent comprising a coenzyme-dependent enzyme and an artificial coenzyme to increase batch size and / or batch homogeneity in the production of diagnostic elements, wherein each of the individual diagnostic elements of a batch substantially are identical.
  • the batch size to a chemical detection reagent comprising a coenzyme-dependent enzyme and a native coenzyme by a factor of 2, preferably by a factor of 3, particularly preferably increased by a factor of 5.
  • Figure 1 Representation of the activity of glucose-dehydrogenase double mutant GlucDH_E96G_E170K (GlucDH-Mut2) on storage of the enzyme in the presence of carbaNAD over a period of 52 weeks at a temperature of 5 ° C and 35 ° C and a relative humidity of 0 % (Desiccant), 75% and 85% respectively.
  • FIG. 2 Representation of the content of carbaNAD (cNAD) during storage of the coenzyme in the presence of glucose dehydrogenase double mutant GlucDH_E96G_E170K over a period of 52 weeks at a temperature of 5 ° C. or 35 ° C. and a relative atmospheric humidity of 0% ( Desiccant), 75% and 85%, respectively.
  • cNAD carbaNAD
  • FIG. 3 Representation of the amino acid sequences of the glucose dehydrogenase double mutants GlucDH_E96G_E170K (GlucDH Mut1) and GlucDH_E170K_K252L (GlucDH Mut2), which were obtained by mutation of wild-type glucose dehydrogenase from Bacillus subtilis.
  • FIG. 4 Representation of the stability of lactate dehydrogenase (LDH) in 2.5% NaCl-containing K / NaP 2 0 7 at pH 8.0 and a temperature of 40 ° C and 50 ° C. Shown is the stability in the presence or absence of the co-factors NAD and carbaNAD (cNAD).
  • LDH lactate dehydrogenase
  • Figure 5 Representation of the stability of glutamate dehydrogenase (GIDH) in 2.5% NaCl-containing K / NaP 2 0 7 at pH 8.0 and a temperature of 40 ° C and 50 ° C. Shown is the stability in the presence or absence of the co-factors NAD and carbaNAD (cNAD).
  • GIDH glutamate dehydrogenase
  • a mixture of glucose dehydrogenase double mutant GlucDH_E170K_K252L and carbaNAD was (a) at a temperature of 5 P C and a relative humidity of 0% (ie in the presence of a desiccant), (b) at a temperature of 5 ° C and a relative Humidity of 75%, (c) stored at a temperature of 35 ° C and a relative humidity of 0%, or (d) at a temperature of 35 ° C and a relative humidity of 85% over a period of 52 weeks ,
  • Lactate dehydrogenase (LDH) was dissolved in 2.5% NaCl-containing K / NaP 2 0 7 solution Temperatures of 40 ° C and 50 ° C exposed. The activity of lactate dehydrogenase was then examined at the beginning and after 3, 21 and 45 hours. The measurement was carried out in the presence and absence of the co-factors NAD and carbaNAD (cNAD).
  • FIG. 4 A graphic representation of the results of these determinations is shown in FIG. 4 of the present application.
  • Glutamate dehydrogenase (GIDH) was exposed in 2.5% NaCl-containing K / NaP 2 0 7 - solution temperatures of 40 ° C and 50 ° C. The activity of glutamate dehydrogenase was then examined at the beginning and after 3, 24 and 45 hours. The measurement was carried out in the presence and absence of the co-factors NAD and carbaNAD (cNAD). These measurements showed that the activity of the enzyme at elevated temperature of 50 ° C in the absence of a co-factor and in the presence of the co-factor NAD strongly decreases, while the stability of the glutamate dehydrogenase in the presence of the co-factor carbaNAD even after several days is still within the range of the initial value.
  • Table 2 The results are summarized in the following Table 2:
  • FIG. 5 of the present application A graphic representation of the results of these determinations is shown in FIG. 5 of the present application.

Abstract

La présente invention concerne un procédé de fabrication d'éléments de diagnostic, un produit de diagnostic comprenant un réactif de détermination chimique stable, ainsi que l'utilisation de ce réactif de détermination chimique stable.
EP14705543.8A 2013-02-22 2014-02-21 Fabrication à haut rendement de bandelettes de test du glucose sanguin Ceased EP2959008A1 (fr)

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EP14705543.8A EP2959008A1 (fr) 2013-02-22 2014-02-21 Fabrication à haut rendement de bandelettes de test du glucose sanguin

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EP13156363.7A EP2770064A1 (fr) 2013-02-22 2013-02-22 Fabrication très efficace de bandes de test de glycémie
PCT/EP2014/053468 WO2014128271A1 (fr) 2013-02-22 2014-02-21 Fabrication à haut rendement de bandelettes de test du glucose sanguin
EP14705543.8A EP2959008A1 (fr) 2013-02-22 2014-02-21 Fabrication à haut rendement de bandelettes de test du glucose sanguin

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EP2959008A1 true EP2959008A1 (fr) 2015-12-30

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EP13156363.7A Ceased EP2770064A1 (fr) 2013-02-22 2013-02-22 Fabrication très efficace de bandes de test de glycémie
EP14705543.8A Ceased EP2959008A1 (fr) 2013-02-22 2014-02-21 Fabrication à haut rendement de bandelettes de test du glucose sanguin

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WO2021139375A1 (fr) * 2020-01-07 2021-07-15 北京九强生物技术股份有限公司 Mutant de glucose-6-phosphate déshydrogénase et utilisation de celui-ci dans la préparation d'un réactif de détection

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US20160010141A1 (en) 2016-01-14
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