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WO1998040739A1 - Biocapteur pour cellules - Google Patents

Biocapteur pour cellules Download PDF

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Publication number
WO1998040739A1
WO1998040739A1 PCT/IL1998/000119 IL9800119W WO9840739A1 WO 1998040739 A1 WO1998040739 A1 WO 1998040739A1 IL 9800119 W IL9800119 W IL 9800119W WO 9840739 A1 WO9840739 A1 WO 9840739A1
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WO
WIPO (PCT)
Prior art keywords
cells
sensing member
specific binding
antibodies
immobilized
Prior art date
Application number
PCT/IL1998/000119
Other languages
English (en)
Inventor
Itamar Willner
Iddo Ben-Dov
Einat Zisman
Shlomo Levi
Original Assignee
Yissum Research Development Company Of The Hebrew University Of Jerusalem
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 Yissum Research Development Company Of The Hebrew University Of Jerusalem filed Critical Yissum Research Development Company Of The Hebrew University Of Jerusalem
Priority to AU66342/98A priority Critical patent/AU6634298A/en
Priority to EP98908267A priority patent/EP0970374A1/fr
Publication of WO1998040739A1 publication Critical patent/WO1998040739A1/fr

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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
    • G01N33/543—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

Definitions

  • the invention is generally in the field of biosensors, and concerns a sensor useful for the determination of the presence, and optionally concentration, of cells in medium, e.g. in urine, blood or mucous.
  • the present invention relates to such sensors, as well as to their use and systems comprising them.
  • Chlamydia trachomatis has been acknowledged as a widespread sexually transmitted microorganism (1). In women, C. trachomatis infections can lead to infertility, ectopic pregnancy, and chronic pelvic pain. In men, C. trachomatis can cause urethritis and epididymitis. Newborns to C. trachomatis infected mothers are in risk of chlamydial conjunctival and pulmonary disease. Cell culture offers a general means for the isolation and identification of C. trachomatis. Various immunoassays for C. trachomatis were also developed, which include enzyme-linked immunosorbant assays (ELISA) and fluorescence labeled antibody assay.
  • ELISA enzyme-linked immunosorbant assays
  • ⁇ f -2.3xl0°f, - ⁇ m / A (1)
  • f 0 is the resonance frequency of the crystal prior to the mass variation
  • A is the surface area of deposited mass.
  • quartz piezoelectric crystals or the so called “quartz crystal microbalance” (QCM) has been adopted for the detection of antigens or antibodies (Ab) in a series of analytical studies. (See review by Suleiman et al, 1994 (2)).
  • QCM immunoassays in solution were disclosed by Roederer (6).
  • the quartz crystal was modified with glycidoxypropyltrime- thoxy silane (GOPS), and the surface-modified crystal was then further modified by anti-human IgG antibody and then applied for the piezoelectric detection of human IgG.
  • the detection limit of the device was determined to be 13 ⁇ g-ml "1 .
  • QCM has been used also to analyze large entities such as microbial cells, using antibody-coated quartz crystals.
  • Candida albicans cells in the concentration range Ixl0 6 -5xl0 8 cells-ml "1 were analyzed by an anti-Candida albicans coated Ab surface (7), E. coli with an anti-E. coli coated surface (8) and protein A-coated crystals acted as piezoelectric sensing interface for various bacteria including Salmonella, Shigella, Yersinia and E. Coli.
  • body fluid samples e.g. urine, serum, plasma, whole blood, cerebro spinal fluid (CSF), amniotic fluid, etc.
  • the present invention makes use of a piezoelectric crystal coated by entities, e.g. monoclonal antibodies, which specifically bind the assayed cells (such entities will be referred to herein at times as "specific binding entities'").
  • entities e.g. monoclonal antibodies
  • a change of mass bound to the crystal reflected by a change of resonance frequency of the crystal is indicative of the presence of said cells in the tested medium.
  • ⁇ / response will be used to denote a change of frequency of the sensor as the result of binding of a mass thereto or release of a mass therefrom.
  • the " ⁇ / response" of the sensor is measured and serves for determination of the presence and optionally the concentration of cells in the assayed liquid medium.
  • a system for assaying cells in a liquid medium comprising a piezoelectric crystal-based sensing member; an electric or electronic unit electrically connectible to the crystal for generating a vibration inducing electric current causing vibrations in said crystal and for measuring its resonance frequency; and a vessel for holding a specimen of said liquid medium and allowing contact thereof with said sensing member; the sensing member comprising a piezoelectric crystal provided with one or more metal plates on its surface, the metal plates having immobilized thereon specific binding entities, which specifically bind to an epitope on the surface of said cells, such that the binding of the cells to the immobilized specific binding entities or release of said cells from the immobilized member bringing to a change of mass of the sensing member, resulting in a change of the sensing member's resonance frequency
  • the assay is performed by first providing conditions allowing binding of the cells to the specified binding entities. Then the change in resonance frequency following binding may be measured. Alternatively, the cells are first allowed to bind to the specific binding entitles and then conditions are provided which cause release of said cells and then the change in frequency following such a release is measured. Regardless of the manner in which the assay is performed, the measured change in resonance frequency serves as an indication for the presence of the assayed cells in the assayed liquid medium and the extent of change serves as an indication of the concentration of the cells in the assayed medium.
  • a method for assaying presence and optionally concentration of cells in a liquid medium comprising:
  • a sensing member comprising a piezoelectric crystal with one or more metal plates on its surface, the metal plates having immobilized thereon specific binding entities which specifically bind to epitopes on the surface of said cells;
  • the specific binding entities are entities which specifically bind to an epitope on the surface of such cells. They may, for example, be antibodies, particularly monoclonal antibodies, which recognize an epitope presented on the surface of said cells. By another example, said entities are lectins which specifically bind to sugar moieties on the surface of said cells.
  • the sensing member may contain on its surface some non-specific binding sites which are capable of adsorption of large molecules, e.g. proteins.
  • macromolecules e.g. proteins
  • Such blocking may be achieved, for example, by soaking the sensing member with bovine serum albumin (BSA), animal (e.g. fish) sera, gelatin, casein, polymers, e.g. polyethyleneglycol, saccharides, e.g. sucrose, trehalose, etc.
  • BSA bovine serum albumin
  • animal e.g. fish
  • polymers e.g. polyethyleneglycol
  • saccharides e.g. sucrose, trehalose, etc.
  • the piezoelectric crystal has to be subjected to alternating electrical field.
  • the piezoelectric crystal used in accordance with the invention is typically a planar crystal having electrodes which are made of an electrically conducting material and which typically have the form of plates (rectangular, circular, etc.), attached to opposite faces of the crystal.
  • the plates carrying the immobilized binding entities can function also as the electrodes which convey the alternating electric field to the crystal.
  • These plates may be selected from a variety of conducting substances, preferred being such which are made from or which are at least coated by a metal or which can chemisorb a sulphur-containing moiety, e.g. gold, platinum or silver.
  • the specific binding entities may be immobilized on the metal plate by means of a linking group having the following general formula (I):
  • Z represents a sulphur-containing moiety which is capable of chemical association with, attachment to or chemisorption onto said metal;
  • R 1 represents a connecting group;
  • Q is a functional group which is capable of forming a covalent bond with a moiety in the specific binding entity.
  • Z may for example be a sulphur atom obtained from a thiol group, a disulfide group, a sulphonate group, or a sulphate group.
  • R 1 may be a covalent bond or may be a peptide or polypeptide or may be selected from a very wide variety of suitable groups such as alkylene, alkenylene, alkynylene phenyl containing chains, and many others. Particular examples of R 1 are a chemical bond or a group having the following formulae (Ha), (lib), (lie) or (lid) A -R 2 -C- -R 2 -NH-
  • R »2 or R may be the same or different and represent straight or branch alkylene, alkenylene, alkynylene having 1-16 carbon atoms or represent a covalent bond, A and B may be the same or different and represent 0 or S,
  • Ph is a phenyl group which is optionally substituted, e.g. by one or more members selected from the group consisting of S0 3 " or alkyl groups.
  • Q may for example be an amine group, capable of binding to a carboxyl residue; a carboxyl group, capable of binding to an amine residue; an isocyanate or isothiocyanate group or an acyl group capable of binding to an amine residue; or a halide group capable of binding to hydroxy residues of title binding entity.
  • Particular examples are the groups -NH 2 ;
  • R and R being, independently a C ⁇ -C 12 alkyl or alkenyl or a phenyl containing chain which is optionally substituted, e.g. by halogen.
  • linking group are those of the following formulae (Tfl) - (IX):
  • n is an integer between 1-6.
  • the degree of change in the resonance frequency correlates with the extent of binding or release of said cells to or from the immobi- lized member and is dependent upon the concentration of said cells in the medium to which the sensor is exposed.
  • the extent of change in the resonance frequencies may be used, in accordance with the invention, as an indication of the concentration of said cells in the medium.
  • sensitivity increasing agents being auxiliary specific binding agents, which are free agents (i.e. a priori non-bound) that can specifically bind to an epitope on the surface of said cells.
  • sensitivity increasing agents may also be conjugated or complexed to large molecules or molecular complexes to further increase the change of mass.
  • secondary auxiliary sensitivity increasing agents may be provided which specifically bind to the first auxiliary sensitivity increasing agents already bound to the cell, thus bringing to an additional mass increase.
  • the senor is coated by a layer comprising more than one specific binding entity, each one being capable of a specific binding to one of the cells to be assayed.
  • the sensor is coated by a layer comprising more than one specific binding entity, each one being capable of a specific binding to one of the cells to be assayed.
  • the sensing member is first contacted with a first cell-specific agent which specifically binds to epitopes on the surface of such cells, and then a change in mass after such contact, measured by a change in the resonance frequency, is an indication that the respective cell is immobilized on the sensing member, and hence existed in the specimen.
  • a second cell-specific binding agent is then contacted with the sensing member and change of resonance frequency serves then as an indication of the presence of the second respective cell in the specimen.
  • use of the sensitivity increasing complexes as described above, may be made.
  • the assayed liquid medium may for example be urine, vaginal mucous, serum, plasma, whole blood, CSF, or generally any body fluid
  • Cells which may be assayed include yeast, e.g. Candida; bacteria, e.g. chlamydia, mycoplasma, niceria monocytogenes and others; as well as viral particles, e.g.
  • Fig. 1 illustrates the sequence of steps in the preparation of a gold (Au) plated piezoelectric crystal to immobilize anti-chlamydia antibodies (Abs) thereto to form a sensor member in accordance with the invention and subsequent use of the sensor for assaying of chlamydia in a medium;
  • Fig. 2 shows time-dependent changes in resonance frequency of sensors, at different concentrations of C trachomatis injected to the assay cell suspended in urine, with the specific binding entity being a mouse-IgG-anti-C trachomatis monoclonal antibody ("Ctrachomatis binding entity”):
  • Fig. 2 A shows results from a sensor where the Ctrachomatis binding entity is bound to Fc-specific, goat-IgG-anti-mouse IgG-Ab immobilized on the sensor;
  • Fig. 2B shows results from a sensor where the Ctrachomatis specific binding entity is bound to an immobilized goat IgG anti-mouse IgG, F(ab') 2 .
  • the concentration of Ctrachomatis are as follows: (a) 0.078 ⁇ g/ml; (b) 0.26 ⁇ g/ml; (c) 0.78 ⁇ g/ml; (d) 2.6 ⁇ g/ml; (e) 7.8 ⁇ g/ml;
  • Fig. 3 illustrates a manner of increasing sensitivity in the determination of chlamydia in a medium;
  • Fig. 4 shows additive changes in resonance frequency of a
  • C.tr achomatis-fxmctionalize ⁇ sensor as a function of time, upon treatment with different antibodies: (a) goat-IgG-anti-mouse IgG (Fc-specific); (b) mouse IgE anti-dinitrophenol; (c) mouse-IgG-anti-dinitrophenol; (d) mouse
  • Fig. 5 shows sequential changes in resonance frequency of a quartz crystal functionalized for detecting Ctrachomatis Ab, as a function of time upon interaction with Ctrachomatis and subsequently with a ti-C trachomatis LPS: (a) Ctrachomatis, 2.6 ⁇ g/ml; (b) anti-C. trachomatis, 0.175 ⁇ g/ml.
  • Fig. 6 Shows time-dependent changes in resonance frequency of a quartz crystal with a sensing interface having immobilized anti-C. trachomatis LPS-Ab attached to goat IgG anti-mouse-IgG, Fc-specific antibodies, pre-treated with 5 mg/ml BSA: (a) addition of a new BSA sample; (b) addition of Ctrachomatis, 0.26 ⁇ g/ml; (c) subsequent to exposure to chlamydial cells as in (b), exposure to anti-C.trachomatis LPS-Ab. 0.175 ⁇ g/ml.
  • Fig. 7 shows time-dependent changes in resonance frequency of a functionalized quartz crystal pre-exposed to urine upon exposure to fresh urine specimens that include variable concentrations of Ctrachomatis: (a) 0.078 ⁇ g/ml; (b) 0.26 ⁇ g/ml; (c) 0.78 ⁇ g/ml; (d) 2.6 ⁇ g/m.
  • Fig. 8 shows time-dependent resonance frequency changes of a functionalized crystal, pre-exposed to urine, upon interaction with fresh urine samples that include variable concentrations of Ctrachomatis and subsequent amplification/confirmation of the primary analysis: (a) exposure of the sensor with Ctrachomatis in urine, 0.26 ⁇ g/ml; (b) subsequent exposure of the sensor of (a) with anti-C.
  • trachomatis LPS-Ab 175 ⁇ g/ml in PBS; (c) exposure of the sensor to Ctrachomatis, 2.6 ⁇ g/ml in urine; (d) subsequent exposure of the sensor of (c) to anti-C. trachomatis LPS- Ab, 0.175 ⁇ g/ml in PBS; (e) exposure of the sensor to a sterile urine sample; (f) subsequent treatment of the sensor of (e) with the anti-C. trachomatis LPS-Ab, 0.175 ⁇ g/ml in PBS.
  • Fig. 9 shows the effect of storage time of the sensors on the resonance frequency changes following exposure to 0.26 ⁇ g/ml.
  • the electrodes were functionalized by mobilization of anti-C. frachomatis antibodies by two different routes: by goat-IgG-anti-mouse-IgG antibodies (filled circles); and by fragmented F(ab') 2 anti-mouse IgG-antibodies (filled squares).
  • Quartz crystals sandwiched between two gold (Au)-electrodes are the preferred sensors of the present invention. All measurements described below were performed using 9 MHz quartz piezocrystals (QPC) (AT cut type) covered with a layer (ca. 0.2 cm 2 ) consisting of sputtered gold (ca. 3000A) on a titanium (Ti) substrate (ca. 500 A) (Seiko EG&G). The frequency measurements were performed using a Quartz Crystal Analyzer (model QCA917, Seiko EG&G) linked to a personal computer.
  • QPC quartz piezocrystals
  • the senor To functionalize the sensor, first it is cleaned by treatment with HCl consisting of a first soaking for two minutes in 1 M HCl solution, then after rinsing and drying, applying 50 ⁇ l concentrated HCL on the gold surfaces for two minutes followed by rinsing.
  • HCl consisting of a first soaking for two minutes in 1 M HCl solution, then after rinsing and drying, applying 50 ⁇ l concentrated HCL on the gold surfaces for two minutes followed by rinsing.
  • the sensing interfaces for sensing Ctrachomatis may be prepared as schematically outlined in Fig. 1: a cystamine monolayer is assembled on the Au-electrodes associated with the quartz crystal by treatment of the crystal, in a test-tube, with a 0.02 M aqueous solution of cystamine dihydrochloride for two hours. The resulting electrode is then rinsed with ethanol and water.
  • PBS phosphate buffer saline
  • the surface densities of the cystamine sub-monolayer and sulfo-SMPB can be determined by following the crystal resonance frequencies at each modification step and the application of the Sauerbrey relation.
  • the resulting functionalized crystal is further modified by linking cysteine residues of anti-mouse IgG Ab or of fragmented F(ab') 2 anti-mouse IgG Ab to the maleimide residues on the monolayer, by two alternative routes: (a) The electrodes are reacted with polyclonal goat IgG anti-mouse IgG, or (b) the electrode are reacted with fragmented polyclonal goat IgG anti-mouse IgG, F(ab') 2 .
  • Table 1 summarizes typical values of the crystal frequency changes at the different steps of modification upon assembly of the 2inti-C.tr achomatis Ab to the anti-mouse IgG-Ab or the F(ab') 2 fragmented anti-mouse IgG-Ab.
  • the surface coverage of the different components are also included in the Table. It should be noted that these frequency changes are only representative values and some deviations can be expected, especially for the primary two layers of cystamine and sulfo-SMPB, due to differences in the roughness of the Au-electrodes.
  • Table 1 typical resonance frequency changes ( ⁇ f) of quartz crystals upon stepwise assembly of the components in the sensing interfaces and the respective calcu-
  • Sensing interface consists of the goat IgG anti-mouse IgG, Fc-specific, Ab as sublayer.
  • Sensing interface consists of the goat F(ab') 2 anti-mouse IgG Ab as sublayer.
  • the resulting sensor units are mounted in an assay cell, and the cell is then ready to be filled with the test solution.
  • the crystal is allowed to stabilize to a constant resonance frequency typically ⁇ 1 Hz. Samples of C trachomatis in PBS of variable concentrations are injected into the cell and the crystal frequency changes can be monitored as a function of time.
  • Fig. 2 shows the frequency changes of crystals tailored by the two methods upon interaction of the electrode with different concentrations of Ctrachomatis. In these experiments, the electrodes are consecutively treated with increased concentrations of C. trachomatis.
  • Fig. 2A shows the resonance frequency changes of a sensor having anti-Ctrachomatis antibody immobilized onto the whole anti-mouse IgG antibody layer.
  • FIG. 2B shows the resonance frequency changes of a sensor having anti-Ctrachomatis sensing layer immobilized onto a fragmented F(ab') 2 anti-mouse IgG layer.
  • the frequency of the crystal decreases, implying the association of Ctrachomatis to the sensing interface.
  • the response-time for the detection of the bacteria is ca. 350 sec. and is defined as the time after which the crystal frequency levels off at low concentrations of Ctrachomatis.
  • sensitivity increasing antibodies can further be increased by secondary sensitivity increasing antibodies which bind to the first sensitivity increasing antibodies.
  • the anti-C. trachomatis antibodies [anti-LPS, anti-P60 or anti-MOMP (major outer membrane protein)] were injected into the assay cell, at 0.175 ⁇ g/ml, and the crystal's resonance frequency changes were followed as a function of time.
  • the anti-C. trachomatis functionalized electrodes were treated, after rinsing, with the polyclonal, Fc-specific, goat IgG-anti-mouse IgG, 0.175 ⁇ g/ml, to obtain further amplification.
  • the crystal frequencies were monitored as a function of time to follow the association of the antibodies.
  • Fig. 4 shows the resonance frequency changes of the sensor after exposure to Ctrachomatis, 2.6 ⁇ g/ml, followed by treatment with a variety of potential sensitivity increasing antibodies: goat anti-mouse IgG (curve a), mouse IgE anti-dinitrophenol (curve b) and mouse IgG anti-dinitrophenol (curve c).
  • FIG. 5 shows the resonance frequency changes of the sensor pre-exposed to the bacteria upon exposure to anti-Ctrachomatis LPS antibodies (curve a), and after subsequent exposure to goat anti-mouse IgG antibodies (curve b).
  • the anti-Ctrachomatis sensing interface is unaffected by foreign proteins such as cytochrome oxidase, cytochrome C or glucose oxidase, it was found that there may occur some non-specific binding to the sensing surface by proteins such as BSA. Interaction of the electrode with BSA at 5 ⁇ g/ml results in a frequency change of ca. -100 Hz due to non-specific adsorption of the protein to the surface.
  • the BSA-treated sensing surface is, however, not further influenced upon interaction with BSA, but reveals the original affinity for Ctrachomatis. Fig.
  • FIG. 6 shows the frequency changes of the crystal pre-treated with BSA upon interaction with a new BSA sample (curve a) and upon treatment with Ctrachomatis 0.26 ⁇ g/ml.
  • ⁇ f -6 Hz
  • the anti-C. trachomatis LPS Ab is associated to the bare antibody layer in the presence of BSA at 5 mg/ml (see Fig. 1).
  • the resulting sensing interface is unaffected in the presence of BSA but reveals analogous activity for sensing C. trachomatis.
  • Fig. 6 (curve c) shows the resonance frequency changes of a BSA pre-treated electrode after-treatment with Ctrachomatis at 0.26 ⁇ g/ml and subsequent incubation with the anti-Ctrachomatis LPS Ab.
  • the observed frequency decrease confirms the primary association of the antigen-bacteria to the sensing interface and amplifies the transduced signal.
  • a preferred embodiment of the invention is aimed at detecting the chlamydia in urine samples.
  • the performance of the sensor in a real urine sample was examined.
  • the BSA-treated sensors were treated with Ctrachomatis free urine. This results in a frequency change of the crystal of ca. -50 Hz due to the non-specific association of the urine ingredients.
  • the resulting probes were insensitive towards additional pure urine specimens but revealed activities in sensing the specific cells in urine samples.
  • Inactivated Ctrachomatis bacteria were dispersed in urine and the crystal was challenged with different cell concentrations in urine.
  • Fig. 7 shows the responses of the sensor in the presence of different samples of urine containing variable concentrations of Chlamydia cells.
  • the crystal frequency decreased as the concentration of the Ctrachomatis in the analyte sample increases. It is noted, however, that the extent of frequency decrease is substantially lower than the values observed when the antigen was dissolved in a pure PBS buffer solution. For example, in the presence of Ctrachomatis at concentrations corresponding to 0.26 and 0.78 mg/ml, the frequencies changes of the electrode in urine are -1 and -9 Hz, whereas in PBS solutions, the frequency changes are -6 and -17 Hz, respectively [cf. Fig. 3(A)]. This implies that the sensitivity of the probe decreases in the presence of urine.
  • Ctrachomatis involves the interaction of the sensing interface with the analyte urine sample followed by rinsing of the assay cell with a PBS buffer solution and then introduction of the anti-LPS antibodies.
  • the sensors having a base layer of whole goat anti-mouse IgG antibodies exhibit a degradation with time, whereas the electrodes where the base antibodies are fragmented F(ab') 2 goat anti-mouse IgG antibody reveal unaltered activity for a period of 90 days of storage.
  • This specific probe may be used in cases where more than one microorganism may be a potential cause of a given infection.
  • Such a probe carries on its surface a plurality of specific binding entities, each one specific for one of the cells to be assayed, e.g. one entity being a specific for Chlamydia and the other being specific for N. Gonorrhaeae.
  • a urine specimen by a specific example, is then contacted with the probe. If both types of cells exist in a specimen, then both such cells will become immobilized on the probe. This will give rise to a change of mass, and by measuring the ⁇ f response indication of binding of cells to the probe will be provided.
  • the probe may be contacted with an agent which specifically binds to the other cell and then a mass change is measured in a similar manner as that described above. If the first step in the above sequence does not give rise to change in mass this may serve as an indication that the cells which became immobilized on the probe were the second type of cells. If there is a change of mass only after the first step in the above sequence and not after the second, this is an indication that only the first type of cells in the specimen. If there is a change of mass after both steps in the sequence, this is an indication that both cells existed in the specimen.

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Abstract

L'invention concerne un système et une méthode qui permettent de détecter la présence et éventuellement de mesurer la concentration de cellules dans un milieu liquide, ainsi qu'un élément de détection destiné à être utilisé avec lesdits système et méthode. L'élément de détection comprend un cristal piézo-électrique disposé à sa surface et comportant une plaque métallique dans laquelle sont immobilisées des entités de fixation spécifiques, qui se fixent spécifiquement sur un épitope porté par la surface desdites cellules. La fixation des cellules sur les entités de fixation spécifiques ou leur libération entraîne une modification de la masse, ce qui modifie la fréquence de résonance du cristal piézo-électrique. En mesurant la modification de la fréquence de résonance, on a une indication sur la présence des cellules testées dans un milieu, et en évaluant l'importance de ladite modification, on peut déterminer la concentration des cellules dans ledit milieu. Dans un mode de réalisation spécifique, les cellules testées sont des Chlamydia et le milieu liquide est l'urine.
PCT/IL1998/000119 1997-03-13 1998-03-13 Biocapteur pour cellules WO1998040739A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU66342/98A AU6634298A (en) 1997-03-13 1998-03-13 Biosensor for cells
EP98908267A EP0970374A1 (fr) 1997-03-13 1998-03-13 Biocapteur pour cellules

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IL12044597A IL120445A0 (en) 1997-03-13 1997-03-13 Biosensor for cells
IL120445 1997-03-13

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Cited By (6)

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WO2001002857A1 (fr) * 1999-04-22 2001-01-11 Akubio Limited Mesure et utilisation des interactions moleculaires
WO2001002858A1 (fr) * 1999-07-05 2001-01-11 Institute Of Molecular Agrobiology Methode de test d'immunodiagnostic pour maladie veterinaire
WO2005050164A3 (fr) * 2003-11-13 2005-08-11 Georgia Tech Res Inst Systemes et procedes de detection
US7171844B2 (en) 2000-04-05 2007-02-06 The Charles Stark Draper Laboratory, Inc. Apparatus and method for measuring the mass of a substance
US7763475B2 (en) 1999-04-22 2010-07-27 Inverness Medical Switzerland Gmbh Measurement and use of molecular interactions
WO2017042262A1 (fr) 2015-09-11 2017-03-16 Bpc Arnold Gmbh Biopharma Consulting Puce de biocapteur

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EP0451687A2 (fr) * 1990-04-12 1991-10-16 Miles Inc. Essai immunologique du type sandwich partiel pour détermination de chlamydia
US5135852A (en) * 1989-07-25 1992-08-04 E. I. Du Pont De Nemours And Company Piezoelectric cell growth biosensing method using polymer-metabolic product complex interactions
WO1994024561A1 (fr) * 1993-04-19 1994-10-27 Kurt Nilsson Biodettecteur contenant un glucide immobilise
WO1995032427A1 (fr) * 1994-05-20 1995-11-30 The Cooper Union For The Advancement Of Science And Art Capteur et procede pour la detection d'especes chimiques predeterminees dans une solution
US5516638A (en) * 1992-11-18 1996-05-14 Calypte, Inc. Immunoassays for the detection of antibodies to Chlamydia trachomatisi in the urine.
WO1997004314A2 (fr) * 1995-07-21 1997-02-06 Yissum Research Development Company Of The Hebrew University Of Jerusalem Determination d'un analyte dans un milieu liquide

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Publication number Priority date Publication date Assignee Title
US4735906A (en) * 1984-11-28 1988-04-05 Texas A&M University Sensor having piezoelectric crystal for microgravimetric immunoassays
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WO2017042262A1 (fr) 2015-09-11 2017-03-16 Bpc Arnold Gmbh Biopharma Consulting Puce de biocapteur
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