EP1397680A1 - Procede et essai chimique par transduction de pression - Google Patents

Procede et essai chimique par transduction de pression

Info

Publication number
EP1397680A1
EP1397680A1 EP02725706A EP02725706A EP1397680A1 EP 1397680 A1 EP1397680 A1 EP 1397680A1 EP 02725706 A EP02725706 A EP 02725706A EP 02725706 A EP02725706 A EP 02725706A EP 1397680 A1 EP1397680 A1 EP 1397680A1
Authority
EP
European Patent Office
Prior art keywords
assay system
analyte
chamber
antibody
test strip
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.)
Withdrawn
Application number
EP02725706A
Other languages
German (de)
English (en)
Other versions
EP1397680A4 (fr
Inventor
Theodore c/o Disan Inc. SAND
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.)
Disan Inc
Original Assignee
Disan Inc
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 Disan Inc filed Critical Disan Inc
Publication of EP1397680A1 publication Critical patent/EP1397680A1/fr
Publication of EP1397680A4 publication Critical patent/EP1397680A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N7/00—Analysing materials by measuring the pressure or volume of a gas or vapour
    • G01N7/14—Analysing materials by measuring the pressure or volume of a gas or vapour by allowing the material to emit a gas or vapour, e.g. water vapour, and measuring a pressure or volume difference
    • G01N7/18—Analysing materials by measuring the pressure or volume of a gas or vapour by allowing the material to emit a gas or vapour, e.g. water vapour, and measuring a pressure or volume difference by allowing the material to react
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00108—Test strips, e.g. paper
    • 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
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/44—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from protozoa
    • G01N2333/455—Eimeria

Definitions

  • the field of the invention is test systems for analytes.
  • Analysis of an analyte in a sample generally includes detection of the analyte.
  • Detection may be classified as either direct or indirect.
  • Direct detection typically includes observations that are readily apparent without any further chemical reaction.
  • Indirect detection on the other hand, often requires a chemical reaction before becoming readily apparent.
  • Indirect detection may include reactions that cause changes in mass. Devices that detect changes in mass, however, are often not amenable to certain types of samples. Another indirect detection involves events measured using labels that are radioisotopes. Such techniques, however, raise safety concerns.
  • Indirect detection may also include reactions that cause a change in color, fluorescence, and luminescence. Such changes, however, may be so slight that they are not readily detectable by the human eye. Slight changes in color of many known systems may be due to relatively small concentrations of analytes present in the samples. In addition, color changes and other photometric characteristics may be spread over too large an area to be very useful. Although devices and chemical reactions may be used to amplify signals, noise is often amplified as well.
  • U.S. Patent 6287851 issued to Delwiche et al. (September 2001) teaches a sensor comprising a chamber having an inlet adapted to admit a liquid sample which gets pumped into a reaction cell using a pump. Problems exist in this system generally because it requires that a liquid sample enter the reaction cell. First, a liquid sample may not always be available. Second a sufficient volume of liquid may not be available to enable pumping to occur. Third, the existence of a pump and porous membrane to separate the liquid portion from the gaseous portion adds complexity to the sensor design.
  • the present invention is directed toward analyzing an analyte contained on a collection strip.
  • the analyte generally reacts with a reagent in a chamber.
  • the reaction evolves a gas.
  • a piezoelectric crystal, polymer or other material that produces an electrical signal in response to pressure changes caused by the gas is in gaseous communication with the chamber and the reaction.
  • Figure 1 is a schematic vertical cross section of a further exemplary assay system.
  • Figure 2 is a schematic view of an exemplary test strip.
  • Figure 3 is a schematic of an exemplary "sandwich" configuration in an exemplary assay system.
  • an assay system 100 generally comprises a base 110 and a housing 120, which forms together with lid 122, a chamber 130. Disposed within the chamber 130 is a strip holder 140 that receives a strip (not shown). Chamber 130 is in gaseous communication with the piezoelectric crystal 160 via gas conduit 150. The piezoelectric crystal 160 produces an electric signal that is processed in a circuitry (not shown) comprising a microprocessor 172 on the circuit board 170.
  • a test strip generally receives a sample comprising an analyte.
  • analyte refers to the at least one substance of which the presence or concentration is be determined. Therefore, contemplated analytes include single molecules, homo-and heterodimers, -trimers and -multimers, molecular assemblies (e.g., K-ATPase, microtubuli, etc.), molecular superstructures (e.g., biological membranes), and various organisms.
  • Particularly preferred analytes include peptides of various molecular weights, nucleic acids (e.g., RNA, DNA, PNA, etc.), carbohydrates, hormones, pharmaceutical agents, lipids, bacteria, viruses, viroids, mammalian cells, parasites, chelated metal ions, or haptenic organic molecules.
  • nucleic acids e.g., RNA, DNA, PNA, etc.
  • carbohydrates hormones, pharmaceutical agents, lipids, bacteria, viruses, viroids, mammalian cells, parasites, chelated metal ions, or haptenic organic molecules.
  • analyte particularly includes complexes of the aforementioned substances and structures with a first compound, wherein the complex formation may be due to a covalent bond, a non-covalent bond (e.g., electrostatic, ionic, hydrophobic interaction, etc.), or any reasonable combination thereof.
  • Particularly contemplated first compounds include various peptides, antibodies or their fragments, nucleic acids, enzyme, and metal ion chelators.
  • contemplated first compounds may further include an enzyme that produces a gas when the enzyme reacts with the reagent. There are numerous methods of coupling an enzyme to a substrate, and all known methods are contemplated suitable for use in conjunction with the teachings herein.
  • enzymes that catalyze a reaction that produces a gaseous product are suitable, however, especially preferred enzymes include peroxidases, ureases, carbonic anhydrases and catalases from various sources. Further contemplated enzymes include decarboxylases (and particularly amino acid decarboxylases, oxalate DC, pyruvate DC, etc.), various dehydrogenases (pyruvate DH, isocitrate DH), and oxidases. Moreover, it should be recognized that more than one enzyme may be employed in the generation of a gas. For example, an enzyme coupled to the first compound may produce a first product, which is then substrate for one or more subsequent enzymatic and/or non-enzymatic reactions in which a gas is formed.
  • a chamber 130 receives a test strip, and the chamber is preferably sealed during the reaction such that the electric signal is sufficient to provide a qualitative assessment of the analyte (e.g., partially sealed).
  • the chamber may be sealed during the reaction such that the electric signal is sufficient to provide a quantitative assessment of the analyte (e.g., hermetically sealed).
  • the volume of the chamber relative to the sample allows a relatively rapid increase in pressure where the chamber is at least partially sealed. Consequently, it is preferred that the chamber volume is no greater than 5 times, and more preferably no greater than 3 times the volume of the sample that is applied to the test strip. It should be appreciated that the volume of the chamber relative to the sample is not meant to be a limitation, and as such the chamber volume may be 20, 50, or even 100 times or more than that of the sample.
  • a piezoelectric crystal 160, or other suitable material is in gaseous communication with the chamber 130. The piezoelectric crystal 160 generates an electrical signal in response to a pressure change in the chamber caused by a reaction between the analyte and a reagent.
  • reaction refers to any substance or substrate that in a chemical reaction (preferably as substrate or cosubstrate of contemplated enzymes) will produce a gaseous compound.
  • gaseous compound as used herein includes all compounds that have in their isolated form at atmospheric pressure a boiling point of lower than 25°C.
  • contemplated substrates may vary considerably. However, especially preferred substrates include H 2 O 2 , urea, HCO 3 " , various amino acids, glucose, ethanol, salicylate, etc.
  • a microprocessor 172 is preferably located on a circuit board 170.
  • a microprocessor may be any appropriate microprocessor (i.e., Intel Pentium 4, AMD Athlon, and so on). Depending on the particular configuration and point of use, contemplated configurations may include a microprocessor that transforms the electrical signal from the piezoelectric material into a user readable output, wherein the microprocessor may or may not be at least partially enclosed in the chamber.
  • a sample is mixed with a solution comprising an antibody that binds the analyte with relatively high affinity (i.e., K D ⁇ 10 6 M ' ') to form a complex, wherein the antibody is further coupled to catalase.
  • the resulting mix is applied to the sample receiving area of the test strip and moves (predominantly via capillary action) towards the wick on the opposite end of the test strip.
  • the screening area on the test strip comprises immobilized analyte in a concentration effective to remove substantially all of the antibody that did not bind to the analyte in the sample (or excess antibody).
  • the capture zone on the strip comprises immobilized antibodies that bind the complexes with relatively high affinity.
  • the amount of bound complexes (bearing a gas producing enzyme) in the capture zone is substantially identical with the amount of analyte.
  • the test strip has a configuration as depicted in Figure 2.
  • the test strip 200 is manufactured on a base of MylarTM (not shown) onto which (from left to right) a sample receiving zone 210 is juxtaposed to an area of nitrocellulose 220, which is adjacent to a screening zone 230.
  • the screening zone 230 is followed by another nitrocellulose area 220 juxtaposed to a capture zone 240, a nitrocellulose area 220, and a wick 250.
  • a sample applied to the sample receiving zone 210 will travel through the nitrocellulose area via the screening zone and capture zone to the wick by virtue of capillary action.
  • a "sandwich format" e.g., a carrier material with capture antibodies binds the antigen, which binds in turn a detector antibody
  • a target antigen e.g., particularly contemplated carriers include nitrocellulose membranes (with optional backing such as a sticky MylarTM membrane) or other membranes to which antibodies can be coupled. It is further preferred that suitable membranes are blocked after application of the capture reagent to minimize non-specific binding of subsequent reagents and/or sample. The sample containing the antigen can then be applied to the capture zone (e.g., via capillary action or pipetting) and incubated for an appropriate amount of time.
  • an exemplary sandwich could have a configuration as depicted in Fig. 3.
  • the sandwich 300 comprises a backing 310 onto which a nitrocellulose membrane 320 is attached. Bound to the membrane 320 is a plurality of capture antibodies 330 that bind the analyte 340. The analyte is detected with a detection antibody 350, which is coupled to an enzyme. Examples
  • a lateral-flow assay was constructed based on commercially available reagents and enzyme conjugates.
  • the assay was developed with a screening zone and a capture zone, in order to generate signals that were directly proportional to the concentration of the test substrate atrazine.
  • This assay was evaluated in a pressure sensing test bed for the ability to respond to atrazine in spiked buffer samples.
  • the test system produced signal differentials of approximately 3.5-fold for samples with 1 ppm atrazine compared to negative samples.
  • the antibody against atrazine was obtained from Biostride, Inc. (Palo Alto, CA). Atrazine-BSA also was obtained from Biostride.
  • the Goat anti-Rabbit IgG was obtained from Jackson ImrnunoResearch Laboratories (West Grove, PA). All other chemicals were obtained from Sigma Chemical, Co. (St. Louis, MO), Aldrich (St. Louis, MO), Pierce Chemical Co. (Rockville, IL) or Pharmacia (Piscataway, NJ).
  • the pressure chamber, the assay strip holder and the mounted pressure sensor were fabricated by J M Speciality Parts (San Diego, CA).
  • the piezofilm was obtained from AMP, Inc (Folsom, CA).
  • the pressure sensors were obtained from SenSym, Inc. (Sunnyvale, CA).
  • Urease, carbonic anhydrase, catalase, peroxidase, and their respective substrates were purchased from Sigma. The enzymes were purified and biotinylated following standard protocols known in the art. Similarly, anti-atrazine antibodies were purchased from Sigma and conjugated with streptavidin following standard protocols known in the art.
  • Streptavidin-containing antibodies and biotin-containing enzymes ware conjugated by mixing (with excess of biotinylated enzyme). The mixture is incubated at room temperature for 2 hr and then overnight at 4°C with gentle shaking. After incubation, the mixture is run on a Sepharose S300 column and the leading edge of the first peak is collected. The collected fractions were evaluated for the presence of antibody reactive with atrazine in an ELISA under standard conditions. The results from ELISA analysis indicated that antibody and enzyme had formed conjugates, since antibody activity was found in the pools made from the leading edge of the elution profiles of both urease and catalase.
  • Nitrocellulose is lightly marked with pencil to indicate the location of the screening and capture zones.
  • the screening zone material is diluted in 10 mM Tris (pH 8.5) to give a concentration of 1 mg/ml. Approximately 10 Jl of the solution is spotted in the appropriate zone.
  • the antibody used for the capture zone is diluted in 10 mM Tris (pH 8.5) to give a concentration of 1 mg/ml. Approximately 8 Jl of the solution is spotted in the appropriate zone.
  • the nitrocellulose is placed in excess 10 mM Tris buffer with 10 mg/ml BSA. The nitrocellulose is blocked for 2 hrs at room temperature, after which it is blotted with paper towels and placed at 45°C for 1 hr.
  • the sample pad material is blocked and dried following known procedures.
  • a strip of sticky mylar (2 mil thickness) is placed on a flat surface.
  • a blocked strip of nitrocellulose is placed on the mylar, leaving some sticky surface at both ends of the strip.
  • the sample pad is placed to the left of the screening zone and the absorbent pad is placed at the other end.
  • the materials are pressed gently into the mylar strip.
  • the completed strips are stored in a desiccator at room temperature.
  • a strip is removed from the desiccator and placed on a flat surface.
  • the antibody- catalase conjugate (at the appropriate dilution made in 10 mM Tris [pH 8.5] with 1 mg/ml BSA) is added to the strip to the left of the screening zone. Approximately 250 Jl of the Tris/BSA solution is added to the sample pad. The liquid is allowed to wick to the sample absorbent pad for 20 - 30 min, depending on the strip's wicking rate. The capture zone and the screening zone are cut from the strip and tested in the pressure sensing test bed. Atrazine was spiked into the antibody-catalase conjugate at the appropriate level and incubated at room temperature for a minimum of 2 hrs before testing.
  • test bed is turned on and allowed to stabilize for 10 min prior to use.
  • the test chamber is cleaned with dH 2 O and 70 % Isopropyl alcohol. 300 Jl of the enzyme substrate solution is added to the chamber.
  • the chamber is sealed and the tubing is connected to the sensor test bed. A timer is started and the initial pressure reading (in mV) is recorded.
  • the chamber is rotated gently for 15 sec, and is rotated for 30 sec at 1 min and again at 3 min. A reading is taken at 5 min.
  • the average (O 1 -0 standard deviation) change in pressure was determined in millivolts produced during a 5 min testing period.
  • the obtained data confirm that contemplated pressure-based assays are responsive to concentrations of the analyste in a sample with a sensitivity of lower limit of detection in the order of 10 ppb at a signal differential of approximately 3.5-fold.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention porte sur un système d'essai comportant une chambre qui accueille une bande test sur laquelle un échantillon comprenant un analyte a été placé. La chambre est en communication gazeuse avec un matériau piézo-électrique qui génère un signal électrique en réaction à une variation de pression dans la chambre due à une réaction entre l'analyte et le réactif.
EP02725706A 2001-05-30 2002-04-15 Procede et essai chimique par transduction de pression Withdrawn EP1397680A4 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US29486101P 2001-05-30 2001-05-30
US294861P 2001-05-30
PCT/US2002/012052 WO2002099430A1 (fr) 2001-05-30 2002-04-15 Procede et essai chimique par transduction de pression

Publications (2)

Publication Number Publication Date
EP1397680A1 true EP1397680A1 (fr) 2004-03-17
EP1397680A4 EP1397680A4 (fr) 2009-06-24

Family

ID=23135261

Family Applications (1)

Application Number Title Priority Date Filing Date
EP02725706A Withdrawn EP1397680A4 (fr) 2001-05-30 2002-04-15 Procede et essai chimique par transduction de pression

Country Status (3)

Country Link
EP (1) EP1397680A4 (fr)
CA (1) CA2448762A1 (fr)
WO (1) WO2002099430A1 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2926815A1 (fr) 2014-04-03 2015-10-07 Institut Curie Dérivés de céphalosporines pour traiter le cancer
CN103994946A (zh) * 2014-06-09 2014-08-20 厦门大学 基于气压检测的多种靶标的高灵敏定量分析方法
CN108918341A (zh) * 2018-07-11 2018-11-30 东莞市松研智达工业设计有限公司 一种用于检测血清或血浆中二氧化碳含量的装置

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3973912A (en) * 1975-06-02 1976-08-10 American Hospital Supply Corporation Micro-gasometric method and apparatus
DE4034174A1 (de) * 1989-10-26 1991-05-02 Ceskoslovenska Akademie Ved Verfahren und vorrichtung zur kontinuierlichen bestimmung des sauerstoffverbrauchs und der kohlendioxidbildung in abgeschlossenen reaktionssystemen
WO1995005592A1 (fr) * 1993-08-19 1995-02-23 Despatch Industries, Inc. Procedes et moyens de test de concentrations en peroxyde dans des agents sterilisants
WO1998028613A1 (fr) * 1996-12-20 1998-07-02 The Regents Of The University Of California Biocapteur d'uree conçu pour la surveillance de l'hemodialyse

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5518895A (en) * 1990-02-15 1996-05-21 Akzo N.V. Device for detecting microorganisms using piezoelectric means
WO1996027432A1 (fr) * 1995-03-07 1996-09-12 Biomolecular Assays, Inc. Reacteur a cycle de pression
WO1999028735A1 (fr) * 1997-12-02 1999-06-10 Smith Allan L Appareil et procede de mesure en simultane des changements de debit massique et de flux thermique
US6287851B1 (en) * 1999-07-09 2001-09-11 The Regents Of The University Of California Sensor for analyzing components of fluids

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3973912A (en) * 1975-06-02 1976-08-10 American Hospital Supply Corporation Micro-gasometric method and apparatus
DE4034174A1 (de) * 1989-10-26 1991-05-02 Ceskoslovenska Akademie Ved Verfahren und vorrichtung zur kontinuierlichen bestimmung des sauerstoffverbrauchs und der kohlendioxidbildung in abgeschlossenen reaktionssystemen
WO1995005592A1 (fr) * 1993-08-19 1995-02-23 Despatch Industries, Inc. Procedes et moyens de test de concentrations en peroxyde dans des agents sterilisants
WO1998028613A1 (fr) * 1996-12-20 1998-07-02 The Regents Of The University Of California Biocapteur d'uree conçu pour la surveillance de l'hemodialyse

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO02099430A1 *

Also Published As

Publication number Publication date
WO2002099430A1 (fr) 2002-12-12
EP1397680A4 (fr) 2009-06-24
CA2448762A1 (fr) 2002-12-12

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