Home - News ( United States | United Kingdom | Italy | Germany ) - Football scores

Showing content from https://patents.google.com/patent/CA2370745A1/en below:

CA2370745A1 - Method for representing biologically activated inductance-altering particles and device for carrying out the method

Publication number

CA2370745A1

CA2370745A1 CA002370745A CA2370745A CA2370745A1 CA 2370745 A1 CA2370745 A1 CA 2370745A1 CA 002370745 A CA002370745 A CA 002370745A CA 2370745 A CA2370745 A CA 2370745A CA 2370745 A1 CA2370745 A1 CA 2370745A1

Authority

CA

Canada

Prior art keywords

particles

inductance

altering

set forth

coated

Prior art date

1999-02-17

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.)

Abandoned

• Espacenet
• Global Dossier
• CIPO
• Discuss

• 239000002245 particle Substances 0.000 title claims abstract description 57
• 238000000034 method Methods 0.000 title claims abstract description 27
• 239000002184 metal Substances 0.000 claims abstract description 22
• 230000005294 ferromagnetic effect Effects 0.000 claims abstract description 18
• 241000700605 Viruses Species 0.000 claims abstract description 9
• 230000010355 oscillation Effects 0.000 claims abstract description 7
• 108090000623 proteins and genes Proteins 0.000 claims abstract description 7
• 238000004062 sedimentation Methods 0.000 claims abstract description 6
• 102000004169 proteins and genes Human genes 0.000 claims abstract description 3
• 239000004809 Teflon Substances 0.000 claims description 8
• 229920006362 Teflon® Polymers 0.000 claims description 8
• 230000004075 alteration Effects 0.000 claims description 6
• 239000003990 capacitor Substances 0.000 claims description 6
• 238000001514 detection method Methods 0.000 claims description 5
• 230000000717 retained effect Effects 0.000 claims description 5
• 125000006850 spacer group Chemical group 0.000 claims description 4
• 239000004033 plastic Substances 0.000 claims description 2
• 238000011144 upstream manufacturing Methods 0.000 claims description 2
• 230000008685 targeting Effects 0.000 claims 2
• 108091034117 Oligonucleotide Proteins 0.000 claims 1
• 239000000463 material Substances 0.000 claims 1
• 239000011859 microparticle Substances 0.000 abstract description 4
• 239000007788 liquid Substances 0.000 abstract description 2
• 241000894006 Bacteria Species 0.000 description 11
• 238000005259 measurement Methods 0.000 description 8
• XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
• 239000000725 suspension Substances 0.000 description 4
• 210000004027 cell Anatomy 0.000 description 3
• 230000005291 magnetic effect Effects 0.000 description 3
• 241000588724 Escherichia coli Species 0.000 description 2
• 239000007864 aqueous solution Substances 0.000 description 2
• 239000011324 bead Substances 0.000 description 2
• 210000000601 blood cell Anatomy 0.000 description 2
• 238000006243 chemical reaction Methods 0.000 description 2
• 239000000470 constituent Substances 0.000 description 2
• 230000008878 coupling Effects 0.000 description 2
• 238000010168 coupling process Methods 0.000 description 2
• 238000005859 coupling reaction Methods 0.000 description 2
• 230000003287 optical effect Effects 0.000 description 2
• 244000052616 bacterial pathogen Species 0.000 description 1
• 239000003153 chemical reaction reagent Substances 0.000 description 1
• 238000011161 development Methods 0.000 description 1
• 230000018109 developmental process Effects 0.000 description 1
• 239000003814 drug Substances 0.000 description 1
• 238000000684 flow cytometry Methods 0.000 description 1
• 230000001900 immune effect Effects 0.000 description 1
• 230000001939 inductive effect Effects 0.000 description 1
• 239000006249 magnetic particle Substances 0.000 description 1
• 244000005700 microbiome Species 0.000 description 1
• 238000003032 molecular docking Methods 0.000 description 1
• 230000001717 pathogenic effect Effects 0.000 description 1
• 230000035699 permeability Effects 0.000 description 1
• 238000001179 sorption measurement Methods 0.000 description 1
• 239000000126 substance Substances 0.000 description 1

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/10—Investigating individual particles
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/04—Investigating sedimentation of particle suspensions
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/10—Investigating individual particles
  • G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
• • 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
  • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
  • G01N33/54326—Magnetic particles
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/06—Investigating concentration of particle suspensions
  • G01N15/0656—Investigating concentration of particle suspensions using electric, e.g. electrostatic methods or magnetic methods
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
  • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
  • G01N2015/019—Biological contaminants; Fouling

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

According to the inventive method for representing biologically activated inductance-altering particles, especially ferromagnetic or superparamagnetic particles, monovalent primary antibodies are mixed with inductance-altering particles in excess, the latter being coated with secondary antibodies.
Aggregated particles are then separated by partial sedimentation, said aggregated particles consisting of a monovalent primary antibody and antibody-coated inductance-altering partial particles. According to a further method, viruses are mixed with ferromagnetic particles in excess, the latter being coated with antibodies that target the sheathing proteins of the viruses, and aggregated particles are separated by partial sedimentation, said aggregated particles consisting of a virus and antibody-coated inductance-altering partial particles. A detecting and counting device for suspended biological microparticles in liquid samples has a delivery line (16) for a sample to be measured which is configured as a measuring line (34) and surrounded by a metal coil which is configured as a measuring coil (36a). The measuring coil is connected to a device (46) for exciting oscillation and measuring resonance events. The metal coil (36a) is placed around a core (50) which is bent approximately into a C shape and which has a gap (52) through which the measuring line (34) is guided.

DESCRIPTION
Method of representing biologically activated inductance-altering particles and device for carr~ring out the method The invention concerns a method of representing biologically activated inductance-altering - in particular ferromagnetic or superparamagnetic - particles. The invention further concerns a device for detecting and counting suspended biological microparticles in liquid samples, in particular for carrying out the specified method.
Hitherto the procedure involved in counting bacteria, blood cells or cell constituents in aqueous solutions has been effected by means of through-flow cytometers or Coulter counters. Here the corresponding particles are colored and identified on the basis of optical signals or 1o counted by capacitive measurement procedures.
In consideration of those factors the inventor set himself the aim of simplifying such measurement operations.
That object is attained by the teaching of the independent claim;
the appendant claims set forth advantageous developments. In addition i5 the scope of the invention embraces all combinations comprising at least two of the features disclosed in the description, the drawing and/or the claims.
In accordance with the invention monovalent primary antibodies are mixed with inductance-altering, in particular ferromagnetic or 20 superparamagnetic, particles in multiple excess, which are coated with secondary antibodies; aggregated particles which comprise a monovalent primary antibody and antibody-coated ferromagnetic partial particles are then separated by means of partial sedimentation in a centrifuge. Instead of primary antibodies it is also possible to use viruses or gene samples, i whose sheathing proteins or spacer molecules are targeted by the secondary antibodies.
In accordance with a further feature of the invention the detecting or counting biological particles are immunologically, phagologically or molecular-biologically joined to aggregated particles which, when subsequently flowing through a metal coil - in particular the gap of a C-shaped metal coil with a ferromagnetic core - trigger measurable and countable alterations in inductance.
It has also proven to be advantageous for inductance-altering 1o particles, before flowing through the metal coil, to be retained by means of an electromagnet in a plastic capillary and there to be joined to the biological particles flowing into the capillary, while the sample in which same were contained is taken out of the capillary. In addition, countable alterations in the natural oscillation frequency are to be produced by the ~5 metal coil as part of an electronic resonant circuit.
In order to obviate the apparatus expenditure in regard to optical measurement and to achieve a higher degree of specificity in comparison with capacitive measurement, a different measurement principle is therefore used for detection of the individual particle: measurement of the 20 alteration in inductance of a microcoil of metal. As however biological particles have a permeability constant a of approximately 1, they have to be previously marked by means of inductance-altering substances for detection and counting procedures by means of a coil. That marking is effected by immunological, phagological or molecular-biological coupling of 25 ferromagnetic or superparamagnetic particles which are monovalently joined either to antibodies, virus docking molecules or gene samples at spacer molecules.
The scope of the invention includes a device of the kind set forth above, having a delivery line for a sample to be measured, which is 3o surrounded as a measuring line by a metal coil as a measuring coil which ' CA 02370745 2001-10-17 in turn is connected to a device for exciting oscillation and measuring resonance events.
In a particular embodiment that metal coil is laid around a core which is bent approximately into a C-shape and whose ends delimit a gap;
the measuring line is laid through that gap.
In accordance with a further feature of the invention the delivery line is connected to a device with capillaries - in particular with Teflon capillaries - ; the latter are associated with an electromagnet and can be arranged in a space surrounded by a pole piece.
to Advantageously provided between the electromagnets and a valve of the delivery line is a branch line for excess sample. In addition at least one resistor and a capacitor can be arranged in front of each device for exciting the oscillations and measuring resonance events, towards the metal coil.
The measuring coil, a piezoelectric pump arranged upstream thereof and a downstream-arranged resistor and capacitor respectively are to be parts of a microsystem-technical unit in accordance with the invention.
Therefore coupling of the ferromagnetic markers occurs in the device which at the same time permits enrichment of the particles to be 2o counted: the markers are retained in the Teflon capillary by means of an electromagnet as a sorption layer, until the entire sample has been pumped into the capillary and at the same time the excess sample has run out of the capillary. Thereupon the magnet is switched off so that the markers freely diffuse and can saturate the surface of the biological particles. The capillary content is then pumped by the above-mentioned piezoelectric pump through the metal coil, in particular through the gap of the metal coil, which is of a C-shaped configuration, with a ferromagnetic core. The metal coil is etched in the form of a spiral onto a circuit board and is connected with capacitor and resistor as a resonant circuit. The resonant circuit is excited by a frequency corresponding to that natural resonant frequency which is generated when an averagely marked biological microparticle is in the gap or the coil. As a result a resonance oscillation always occurs in the resonant circuit when a corresponding microparticle passes through the coil.
An example of the use of that method is the detection of coli bacteria in water samples. For that purpose monovalent primary E.-coli specific antibodies are conjugated with secondary antibodies coupled to magnetic beads. The suspension of those conjugates is pumped into the Teflon capillary and fixed there by means of an electromagnet. When the water sample to be investigated flows through the capillary, coli bacteria 1o are retained to the conjugates by way of the primary antibodies. After the magnet is switched off the suspension of magnetically marked coli bacteria can be pumped through the measuring coil or the gap of the metal coil.
The number of resonance events in the connected resonant circuit corresponds to the number of coli bacteria in the original water sample.
By virtue of the use of that arrangement and the corresponding conjugates, it is possible to automatically count bacteria without the expensive use of through-flow cytometry. Furthermore it is possible with that measuring method to achieve miniaturization of the detection arrangement.
The described procedure is used for detecting and counting particles such as bacteria, cells or cell constituents in aqueous solutions. That procedure permits miniaturization of the automatic particle counting method. For that purpose the particles are marked prior to the measurement procedure by the reaction with monovalent antibody-coated or virus-coated ferromagnetic particles. Inductive measurement is based on passage of the ferromagnetic particles aggregated with the biological particles through the microcoil, designed in the above-described manner, of an electronic resonant circuit. The resonance events which occur upon such particle passage are counted.
The device according to the invention can be used in medicine, microbiology and hygiene, for example for counting out blood cells; it is possible to count out ecologically relevant micro-organisms or detect pathogenic germs.
Further advantages, features and details of the invention will be apparent from the description hereinafter of a preferred embodiment and with reference to the drawing in which:
Figures 1 and 3 each show a diagrammatic view relating to a method according to the invention, and Figure 2 is a diagrammatic perspective view of a detail from Figures 1 and 3.
1o Prior to a method of detecting coli bacteria in a water sample Z
supplied through a line 10 monovalent primary E.-coli-specific antibodies are conjugated to secondary antibodies coupled to magnetic beads. The line for the monovalent magnetic particles F is denoted by reference 12.
Both lines 10, 12 include hose pumps 14 and downstream of same are combined to form a common delivery line 16.
The reagent with ferromagnetic, biologically activated particles is pumped by way of the lines 12 and 16 into a Teflon capillary 20 and is fixed there by means of an electromagnet 22 whose magnetic coil is identified by reference numeral 24 and with which there is associated the 2o Teflon capillary 20 which is wound on in a z-shape, in a concentric pole piece 26. The latter with a pole pin 28 surrounded thereby at a radial spacing defines an annular space 30 for the Teflon capillary.
When the water sample Z to be investigated flows through the capillary 20 coli bacteria as biological particles to be counted are retained 2s by way of the primary antibodies to the ferromagnetic conjugates. After the electromagnet 22 is switched off the suspension of magnetically marked coli bacteria can be transported by virtue of a piezoelectric pump 32 in a measuring line 34 through an etched metal coil as a measuring coil 36 of a microsystem-technical unit 40. The counted particles are 3o discharged therefrom in the direction indicated by the arrow X.

In the embodiment of Figure 3 the suspension is transported in the measuring line 35 through the gap 52 of a ferromagnetic core 50 of a measuring coil 36a, the core 50 being curved in a C-shape.
The free ends 38, 38a of the measuring coil 36, 36a - downstream of a resistor 42 and a capacitor 44 - are connected to a device 46 for exciting the oscillation and for measuring resonance events; there conversion into counting pulses takes place.
The number of resonance events in the connected resonant circuit corresponds to the number of coli bacteria in the original water sample Z.
l0 Provided between the Teflon capillary 20 and the piezoelectric pump 32 is a line branch 18 - which includes a valve 48 - for excess sample portions Q, with a valve 48 being connected downstream thereof in the delivery line 16.

1. A method of representing biologically activated inductance-altering, in particular ferromagnetic or superparamagnetic, particles, characterized in that monovalent primary antibodies are mixed with inductance-altering particles in excess, which are coated with secondary antibodies, and then aggregated particles which comprise a monovalent primary antibody and antibody-coated inductance-altering partial particles are separated by means of partial sedimentation.

2. A method of representing biologically activated inductance-altering, in particular ferromagnetic or superparamagnetic, particles, characterized in that viruses are mixed with inductance-altering particles in excess, which are coated with antibodies targeting the sheathing proteins of the viruses, and then aggregated particles which comprise a virus and antibody-coated inductance-altering partial particles are separated by means of partial sedimentation.

3. A method of representing biologically activated inductance-altering, in particular ferromagnetic or superparamagnetic, particles, characterized in that spacer molecule-coupled oligonucleotide gene samples are mixed with inductance-altering particles in excess, which are coated with antibodies targeting the spacer molecules, and then aggregated particles which comprise a gene sample and antibody-coated inductance-altering partial particles are separated by means of partial sedimentation.

4. A method as set forth in one of claims 1 through 3 characterized in that biological detection or counting particles are immunologically, phagologically or molecular-biologically combined with the aggregated particles which as markers when subsequently flowing through a metal coil trigger off measurable and countable alterations in inductance.

5. A method as set forth in claim 4 characterized in that when flowing through the gap at a core, which is curved substantially in a C-shape, of a metal coil the markers trigger off measurable and countable alterations in inductance.

6. A method as set forth in claim 4 or claim 5 characterized in that inductance-altering particles are retained prior to flowing through the metal coil by means of an electromagnet in a plastic capillary and are combined there with the biological particles flowing into the capillary while the sample containing same is passed out of the capillary.

7. A method as set forth in one of claims 4 through 6 characterized in that countable alterations in the natural resonant frequency are produced by the metal coil as part of an electronic resonant circuit when the inductance-altering particles flow therethrough.

8. A device for carrying out the method as set forth in one of claims 1 - 7, for detecting a biological particle which is conveyed through a conveyor line and which is bonded to a marking particle of inductance-altering, in particular ferromagnetic or superparamagnetic material, wherein the delivery line (16) for a sample to be measured is surrounded as a measuring line (34) by a metal coil as a measuring coil (36, 36a) and same is connected to a device (46) for exciting oscillation and measuring resonance events, and wherein the metal coil (36a) is laid around a core (50) which is curved substantially in a C-shaped configuration and the core has a gap (52) through which the measuring line (34) is passed.

9. A device as set forth in claim 8 characterized in that the marking particle is monovalently bonded to at least one biological particle.

10. A device as set forth in claim 8 or claim 9 characterized in that the delivery line (16) is connected to a device with capillaries (20), in particular Teflon capillaries, and the latter are associated with an electromagnet (22).

11. A device as set forth in claim 10 characterized in that the capillary or capillaries (20) are arranged in a space (30) surrounded by a pole piece (24).

12. A device as set forth in one of claims 8 through 11 characterized in that arranged between the electromagnet (22) and a valve (48) of the delivery line (16) is a branch line (18) for excess samples (Q).

13. A device as set forth in one of claims 8 through 12 characterized in that arranged upstream of the device (46) for exciting the resonance and measuring resonance events towards the metal coil (36, 36a) are at least one resistor (42) and a capacitor (44).

14. A device as set forth in one of claims 8 through 13 characterized in that the measuring coil (36, 36a) with upstream-arranged piezoelectric pump (32) and downstream-arranged resistor (42) and capacitor (44) respectively are parts of a microsystem-technical unit (40).

• 2000
  • 2000-02-15 CA CA002370745A patent/CA2370745A1/en not_active Abandoned
  • 2000-02-15 WO PCT/EP2000/001214 patent/WO2000049407A2/en not_active Application Discontinuation
  • 2000-02-15 AU AU41020/00A patent/AU4102000A/en not_active Abandoned
  • 2000-02-15 EP EP00920436A patent/EP1198712A2/en not_active Withdrawn

RetroSearch is an open source project built by @garambo | Open a GitHub Issue

Search and Browse the WWW like it's 1997 | Search results from DuckDuckGo

HTML: 3.2 | Encoding: UTF-8 | Version: 0.7.4