CA2307280A1 - Analyte assay using particulate labels - Google Patents

Analyte assay using particulate labels Download PDF

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Publication number
CA2307280A1
CA2307280A1 CA002307280A CA2307280A CA2307280A1 CA 2307280 A1 CA2307280 A1 CA 2307280A1 CA 002307280 A CA002307280 A CA 002307280A CA 2307280 A CA2307280 A CA 2307280A CA 2307280 A1 CA2307280 A1 CA 2307280A1
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Prior art keywords
particle
light
particles
scattered
analyte
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CA002307280A
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French (fr)
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CA2307280C (en
Inventor
Juan Yguerabide
Evangelina E. Yguerabide
David E. Kohne
Jeffrey T. Jackson
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University of California
Life Technologies Corp
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Individual
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Priority to CA2719432A priority Critical patent/CA2719432A1/en
Publication of CA2307280A1 publication Critical patent/CA2307280A1/en
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    • 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
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
    • 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
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
    • 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
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1425—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its control arrangement
    • 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
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1456—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • 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
    • G01N2015/0038—Investigating nanoparticles
    • 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
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N2015/1493—Particle size
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/805—Optical property
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/815—Test for named compound or class of compounds
    • Y10S436/817—Steroids or hormones

Abstract

Method for specific detection of one or more analytes in a sample. The method includes specifically associating any one or more analytes in the sample with a scattered-light detectable particle, illuminating any particle associated with the analytes with light under conditions which produce scattered light from the particle and in which light scattered from one or more particles can be detected by a human eye with less than 500 times magnification and without electronic amplification. The method also includes detecting the light scattered by any such particles under those conditions as a measure of the presence of the analytes.

Claims (63)

1. A method for selection of a particle type for detection of an analyte by light scattering, said method comprising the steps of:
a) providing light scattering signal strength versus particle size information for candidate particles; and b) selecting a particle of a size between 1 and 500 nm inclusive which provides acceptable dynamic range, acceptable low concentration detection sensitivity, and acceptable detection resolution based on said information, wherein the acceptable dynamic range, acceptable low concentration detection sensitivity, and acceptable detection resolution are determined by the concentration levels expected for said analyte in samples to be analyzed and are sufficient to allow non-evanescent light scattered from one or more said particles to be detected by a human eye with less than 500 times magnification and without electronic amplification.
2. The method of claim 1, wherein said detection of an analyte by light scattering is performed in a solid-phase assay, and said selecting further comprises identification of a light scattering particle which co-optimizes particle density and particle size, thereby providing a useful combination of signal strength and dynamic range.
3. The method of claim 2, wherein the light scattering signal consists of an integrated scattered light signal.
4. The method of claim 2, wherein the light scattering signal consists of light scattered from an individual particle or a single aggregate containing a plurality of particles.
5. The method of claim 1, wherein said detection of an analyte is performed in a liquid phase assay.
6. The method of claim 5, wherein the light scattering signal consists of an integrated scattered light signal.
7. The method of claim 5, wherein the light scattering signal consists of light scattered from an individual particle or a single aggregate containing a plurality of particles.
8. A method for specific detection of one or more analytes in a sample, comprising the steps of:
specifically associating any said one or more analytes in said sample with a scattered-light detectable particle of a size between 1 and 500 nm inclusive;
illuminating any said particles associated with said analytes with non-evanescent wave light under conditions which produce scattered light from said particle and in which light scattered from one or more said particles can be detected by a human eye with less than 500 times magnification and without electronic amplification; and detecting said light scattered by any said particles under said conditions as a measure of the presence of said one or more analytes, wherein said particles or a multi-particle structure comprising a plurality of said particles is detected singly, wherein said particles are flowed by at least one detector.
9. The method of claim 8, wherein said particles are flowed by said at least one detector in a structure selected from the group consisting of capillaries, microchannels, and spectrophotometric or flow cytometry flow cells.
10. The method of claim 8, wherein said analyte comprises a nucleic acid sequence, and at least one scattered light detectable particle is attached to at least two probe nucleic acid strands, wherein said at least two probe nucleic acid strands bind to different regions of said nucleic acid sequence; and said detecting further comprises flowing said nucleic acid sequences by the detector one at a time and distinguishing single particles from two or more particles, wherein detection of two or more particles is indicative of the presence of said nucleic acid sequence.
11. The method of claim 8, wherein said analyte comprises a cell or a cellular constituent and said detecting further comprises discriminating light scattering of said scattered light detectable particles from light scattering from said cell.
12. The method of claim 11, wherein said discriminating comprises providing two detectors such that the relative levels of cell specific light scattering and particle specific light scattering differ for said two detectors.
13. The method of claim 8, wherein said detecting provides a light scattering signal consisting of light scattered from an individual particle or a single aggregate containing a plurality of particles.
14. The method of claim 8, wherein said detecting provides a light scattering signal the characteristics which comprise at least one of intensity, polarization , wavelength, and angle of observation.
15. The method of claim 8, wherein said detecting provides a light scattering signal indicative of changes in one or more properties of the scattered light detectable properties of individual particles or multi-particle aggregates as said particles or aggregates move in a solution.
16. The method of claim 8, wherein a plurality of different particles with different sizes or compositions or both are distinguishably detected, each said different particle specifically associating with a different analyte.
17. A method for specific detection of one or more analytes in a sample, comprising the steps of:
specifically associating any said one or more analytes in said sample with a scattered-light detectable particle of a size between 1 and 500 nm inclusive;
illuminating any said particles associated with said analytes with non-evanescent wave light under conditions which produce scattered light from said particle and in which light scattered from one or more said particles can be detected by a human eye with less than 500 times magnification and without electronic amplification; and detecting said light scattered by any said particles under said conditions as a measure of the presence of said one or more analytes, wherein said detecting comprises spatially discriminating or classifying or both scattered light from said particles, thereby separately identifying individual particles or mufti-particle structures.
18. The method of claim 17, wherein said discriminating or classifying comprises electronic image processing and analysis.
19. The method of claim 17, wherein said detection of an analyte by light scattering is performed in a solid-phase assay.
20. The method of claim 17, wherein said detection of an analyte is performed in a liquid phase assay.
21. The method of claim 17, wherein the characteristics of the light scattering signal measured comprise at least one of intensity, polarization, wavelength, and angle of observation.
22. The method of claim 17, wherein the light scattering signal measured detects changes in one or more properties of the scattered light detectable properties of individual particles or mufti-particle aggregates as said particles or aggregates move in a solution.
23. The method of claim 17, wherein a plurality of different particles with different sizes or compositions or both are distinguishably detected, each said different particle specifically associating with a different analyte.
24. A method for specific detection of one or more analytes in a sample, comprising the steps of:
specifically associating any said one or more analytes in said sample with a scattered-light detectable particle of a size between 1 and 500 nm inclusive, illuminating any said particles associated with said analytes with non-evanescent wave light under conditions which produce scattered light from said particle and in which light scattered from one or more said particles can be detected by a human eye with less than 500 times magnification and without electronic amplification, and detecting said light scattered by any said particles under said conditions as a measure of the presence of said one or more analytes, wherein said scattered light detectable particles also possess magnetic or electrophoretic properties, and said properties are used to localize said particles during performance of an assay or to separate particles associated with any said one or more analytes from particles not so associated or both.
25. The method of claim 24, wherein said detection of an analyte by light scattering is performed in a solid-phase assay.
26. The method of claim 24, wherein said detection of an analyte is performed in a liquid phase assay.
27. The method of claim 24, wherein the characteristics of the light scattering signal measured comprise at least one of intensity, polarization, wavelength, and angle of observation.
28. The method of claim 24, wherein the light scattering signal measured detects changes in one or more properties of the scattered light detectable properties of individual particles or multi-particle aggregates as said particles or aggregates move in a solution.
29. The method of claim 24, wherein a plurality of different particles with different sizes or compositions or both are distinguishably detected, each said different particle specifically associating with a different analyte.
30. A method for specific detection of one or more nucleic acid analytes comprising target nucleic acid sequences in a sample, comprising the steps of:
specifically associating any of said one or more nucleic acid analytes in said sample with a scattered-light detectable particle of a size between 1 and 500 nm inclusive, wherein said specific associating involves specific hybridization of at least one analyte-specific nucleic acid probe with any of said one or more nucleic acid analytes, wherein said particle is directly attached to a said probe or a said probe has a ligand for attachment of said particle, illuminating any said particles associated with said nucleic acid analytes with non-evanescent wave light under conditions which produce scattered light from said particle and in which light scattered from one or more said particles can be detected by a human eye with less than 500 times magnification and without electronic amplification, and detecting said light scattered by any said particles under said conditions as a measure of the presence of said one or more nucleic acid analytes.
31. The method of claim 30, wherein said detection of an analyte by light scattering is performed in a solid-phase assay.
32. The method of claim 30, wherein said detection of an analyte is performed in a liquid phase assay.
33. The method of claim 30, wherein said one or more nucleic acid analytes comprise a plurality of target nucleic acid sequences;
said plurality of nucleic acid analytes or a plurality of nucleic acid probes are attached in separate spots to a solid surface; and light scattering from one or more of said separate spots is detected as a measure of the presence or amount of at least one of said one or more nucleic acid analytes in said sample.
34. The method of claim 33, wherein determination of an integrated scattered light intensity of a said spot is used as an indicator of the amount of said nucleic acid analyte in said sample.
35. The method of claim 33, wherein the number of said scattered light detectable particles in at least a portion of a said spot is used as an indicator of the amount of a said nucleic acid analyte in said sample.
36. The method of claim 35, wherein the light scattering signals from individual scattered light detectable particles in said at least a portion of a said spot are summed to provide a summed light scattering signal indicative of the amount of a said nucleic acid analyte in said sample.
37. The method of claim 33, wherein light scattering from each said spot is detected by scanning of said separate spots.
38. The method of claim 33, wherein light scattering from each said spot is detected using separate detectors for each said spot.
39. The method of claim 33, wherein light scattering from each said spot is detected using simultaneous imaging of said plurality of spots.
40. The method of claim 39, wherein at least one image of said plurality of spots is analyzed using electronic image processing.
41. The method of claim 33, wherein said plurality of spots comprises an array.
42. The method of claim 41, wherein said array is adapted for sequencing by hybridization or determination of expression level of at least one nucleic acid sequence.
43. The method of claim 30 wherein a plurality of different particles with different sizes or compositions or both are distinguishably detected, each said different particle specifically associating with a different nucleic acid analyte.
44. A method for identifying the presence or amount of at least one cell type or organism in a sample, comprising detecting the presence or amount of at least one scattered light detectable particle of a size between 1 and 500 nm inclusive specifically associated with said at least one cell type or organism:
wherein said at least one scattered light detectable particle is illuminated with non-evanescent wave light under conditions which produce scattered light from said at least one particle and in which light scattered from said at least one particle can be detected by a human eye with less than 500 times magnification and without electronic amplification, and light scattered by any of said at least one particle is detected as a measure of the presence or amount of any of said at least one cell type or organism.
45. The method of claim 44, wherein said identifying the amount of said cell type or organism further comprises determining the amount of cells or organisms with at least one of said particles specifically associated with said cell or organism relative to cells or organisms without said at least one of said particles specifically associated therewith.
46. The method of claim 44, wherein the presence or amount of at least one of a plurality of different yell types or organisms is distinguished or identified using a plurality of different particle types, wherein each of said plurality of different particle types specifically associates with a different cell type or organism, and the light scattering signal from each of said plurality of different particle types is distinguishable from each of the other particle types of said plurality of different particle types.
47. The method of claim 44, wherein the presence or amount of at least one of a plurality of different cell types or organisms is distinguished or identified using a plurality of different particle types, wherein each of said plurality of different particle types specifically associates with at least one of said plurality of different cell types or organisms, and the light scattering signal from each of said plurality of different particle types is distinguishable from each of the other particle types of said plurality of different particle types.
48. The method of claim 44, wherein said determining the presence or amount of at least one cell type or organism comprises identifying said at least one cell type or organism.
49. The method of claim 44, wherein said particle is intracellular.
50. The method of claim 44, wherein a said particle comprises gold or silver or both.
51. A method for detecting the presence or amount of at least one analyte in a cell type or organism, comprising the step of detecting the presence or amount of at least one scattered light detectable particle of a size between 1 and 500 nm inclusive specifically associated with said at least one cell type or organism:
wherein said at least one scattered light detectable particle is illuminated with non-evanescent wave light under conditions which produce scattered light from said at least one particle and in which light scattered from said at least one particle can be detected by a human eye with less than 500 times magnification and without electronic amplification, and light scattered by any of said at least one particle is detected as a measure of the presence or amount of said at least one analyte in a said cell type or organism.
52. The method of claim 51, wherein at least one said analyte is an intracellular protein.
53. The method of claim 51, wherein said at least one analyte is a plurality of analytes, and said at least one scattered light detectable particle is a plurality of distinguishable particles.
54. A method for monitoring a cell or organism, comprising the steps of:
placing at least one scattered light detectable particle of a size between 1 and 500 nm inclusive within a cell; and detecting the presence of at least one said scattered light detectable particle within said cell or organism as an indication of the presence of said cell or organism, wherein said at least one scattered light detectable particle is illuminated with non-evanescent wave light under conditions which produce scattered light from said at least one particle and in which light scattered from said at least one particle can be detected by a human eye with less than 500 times magnification and without electronic amplification.
55. A method for identifying or characterizing a potential pharmaceutical agent, comprising the steps of:
screening a library comprising a plurality of potential targets or a plurality of potential pharmaceutical agents to detect an interaction between a target and a said potential pharmaceutical agent, wherein said screening comprises specifically associating any of said plurality of potential targets or potential pharmaceutical agents or an expression product or mRNA with a scattered-light detectable particle of a size between 1 and 500 nm inclusive, illuminating any said particles associated with any said plurality of potential targets or potential pharmaceutical agents or expression product or mRNA with non-evanescent wave light under conditions which produce scattered light from said particle and in which light scattered from one or more said particles can be detected by a human eye with less than 500 times magnification and without electronic amplification, and detecting said light scattered by any said particles under said conditions such that detection of said light is indicative of a said interaction.
56. The method of claim 55, wherein said interaction comprises a binding interaction between a target or potential target and a said potential pharmaceutical agent.
57. The method of claim 55, wherein said interaction comprises modulation of a drug target system.
58. The method of claim 57, wherein said determination of said modulation comprises determination of the level of expression of a component of said system.
59. The method of claim 55, wherein said detecting said light scattered by any said particles is performed in a homogeneous assay.
60. The method of claim 55, wherein said plurality of potential targets or plurality of potential pharmaceutical agents is in a spatially addressable library.
61. The method of claim 55, wherein each of said plurality of potential pharmaceutical agents is attached to a solid substrate particle.
62. The method of claim 61, wherein a said scattered-light detectable particle also comprise a magnetic or ferro-electric composition, and said method further comprises manipulating the position of said particles associated with a said potential pharmaceutical agent attached to a said solid substrate particle.
63. The method of claim 55, wherein the amount of said expression product or mRNA is altered by said interaction.
CA2307280A 1997-10-17 1998-10-16 Analyte assay using particulate labels Expired - Fee Related CA2307280C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CA2719432A CA2719432A1 (en) 1997-10-17 1998-10-16 Analyte assay using particulate labels

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US08/953,713 US6586193B2 (en) 1996-04-25 1997-10-17 Analyte assay using particulate labels
US08/953,713 1997-10-17
PCT/US1998/023160 WO1999020789A1 (en) 1997-10-17 1998-10-16 Analyte assay using particulate labels

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CA2719432A Division CA2719432A1 (en) 1997-10-17 1998-10-16 Analyte assay using particulate labels

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CA2307280A1 true CA2307280A1 (en) 1999-04-29
CA2307280C CA2307280C (en) 2010-12-14

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US (7) US6586193B2 (en)
EP (1) EP1023456A1 (en)
JP (1) JP4247770B2 (en)
CN (1) CN100379876C (en)
AU (1) AU1294399A (en)
BR (1) BR9814821A (en)
CA (2) CA2719432A1 (en)
HK (1) HK1034291A1 (en)
IL (1) IL135696A (en)
RU (1) RU2217498C2 (en)
WO (1) WO1999020789A1 (en)

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