WO2003091731A1 - Systeme et procede d'analyse multiparametrique de substances a analyser - Google Patents

Systeme et procede d'analyse multiparametrique de substances a analyser Download PDF

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Publication number
WO2003091731A1
WO2003091731A1 PCT/GB2003/001725 GB0301725W WO03091731A1 WO 2003091731 A1 WO2003091731 A1 WO 2003091731A1 GB 0301725 W GB0301725 W GB 0301725W WO 03091731 A1 WO03091731 A1 WO 03091731A1
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WO
WIPO (PCT)
Prior art keywords
analytes
support
analyte
substrate
identification
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PCT/GB2003/001725
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English (en)
Inventor
Christian Bunke
Caroline Garey
Jodie Hadley
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Smartbead Technologies Limited
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.)
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Publication date
Application filed by Smartbead Technologies Limited filed Critical Smartbead Technologies Limited
Priority to AU2003224297A priority Critical patent/AU2003224297A1/en
Publication of WO2003091731A1 publication Critical patent/WO2003091731A1/fr

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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
    • 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

Definitions

  • the present invention relates to a system for multiparameter analysis of analytes; moreover, the invention also concerns a method of performing such multiparameter analysis of analytes.
  • a chemically attached label such as a radionucleotide or fluorescent probe
  • microarray In a United States patent no. US 6, 027, 880, a microarray is described.
  • the microarray concerns an integrated circuit whose two dimensional surface is partitioned into a plurality of spatially disposed sites, each site corresponding to an individual experiment. Each individual experiment is provided with one or more corresponding analyte e.g. nucleotides thereat. Each site is effectively labelled by virtue of its spatial position on the surface of the integrated circuit.
  • This use of microarray technology has become the most common approach of testing large number of oligonucleotides against whole genome samples.
  • a major drawback with microarrays is that they have limitations on the number of probes (query molecules), which can be deposited at the spatial position sites, while maintaining the quality of the experiment results. There are also challenges with high background noise and expensive manufacturing procedures for these microarrays.
  • microarrays for the use in fields such as cancer research, genotyping, neurobiology, toxicology and many others.
  • the number of probes tested on each microarray has increased in recent years to hundreds or even several thousand, the demand for associated manufacturing equipment miniaturization and specialized materials handling has rendered the fabrication of such microarrays increasingly complex and costly.
  • each contemporary microarray is arranged to allow parallel analysis of up to 12000 probes in the form of gene fragments.
  • the characteristics of the probes being monitored on such microarrays must often also be known and isolated beforehand; such prior knowledge makes it a complicated and costly process to manufacture specific microarrays to customer requirements for each different type of organism, species or specific tests to be studied.
  • microarrays Further disadvantages associated with microarrays are low flexibility, poor customisation properties, long manufacturing turnaround times, high cost of reagents and poor sensitivity.
  • Other techniques used to improve the reaction kinetics and therefore the quality of results obtained from microarrays include for example, improvements to surface-to-volume ratios of microarrays. This can include the use of channels and porous materials as described in Akzo Nobel's published international PCT application no. WO 99/02266. Another method of increasing the surface area for the sample attachment is described in the United States patent no.
  • Another approach is to perform simultaneous testing, so called multiplexing, which uses several coded arrays in one vessel rather than a microarray.
  • This can include the use of programmable matrices with memories as described in IRORI's published international PCT application no. WO 96/36436.
  • a recording device stores the information of what molecules and biological materials are linked to the matrix material of each programmable matrice.
  • These matrices can be in solution in one vessel or each linked to a well of a microtitreplate.
  • Several matrices can also be arranged in an array taking the form of a microarray. Other particle array solutions from Virtual Arrays Inc and University of Hertfordshire are described in the published international PCT applications no.
  • WO 00/63419 and WO 00/01475 respectively.
  • These particle based arrays allow greater customisation of the probes (query molecules), which probes are attached to coded particle arrays and tested against a test sample, than the positional based microarray solutions.
  • This solution has problems with cross reactivity of probes (query molecules) when performing many multiplexed assays, each attached to uniquely coded arrays, in one vessel.
  • the reporter systems described previously are used to detect the interaction between probes (query molecules) and the test sample.
  • a first object of the invention is to provide an improved system for the detection of multiparameter characteristics of analytes.
  • a second object of the invention is to improve the parallel testing throughput of currently used two-dimensional arrays systems.
  • the system is of advantage in that it is capable of addressing at least one of the aforementioned objects of the invention.
  • the system is beneficial in that it is flexible and can be used to complement and/or improve existing array technology.
  • the nature of at least one primary analyte is known by virtue of its position of binding on a solid substrate and the nature of at least one secondary analyte is determined by its support's identification means.
  • This provides the information of binding characteristics of primary and secondary analytes by analysing their interaction. As reactions of secondary analytes in the system are tagged by individual identifiable supports, the throughput previously achieved using arrays is efficiently improved. Such improvement also allows the use of adapted conventional reading means rather than requiring new more advanced readers used for reading arrays with greater spotting density. As multiple analytes are present in the system, an improvement in the number of parameters that can be analysed simultaneously is achieved.
  • the identification means is a barcode allowing for easy identification using well-established standards and methods.
  • the analytes comprises a probe and/or a test sample, which are bound to identifiable supports.
  • the identification of the reaction can hence be established by the final position of the support on the substrate and the identification code of the support.
  • the probes are spotted or synthesised onto the main surface of the array allowing the use for commonly used microarrays, which allows easy adaptation of the new system which combines the established microarrays with particle based support assay technology. Further it is preferable that the support with identification means is smaller than the area of spotted or synthesised probe on the microarray surface.
  • At least two primary analytes and at least two secondary analytes are used in an experiment to truly benefit from the multiparameter analysis of the solution.
  • the method is of advantage in that it is capable of addressing at least one of the aforementioned objects of the invention.
  • Figure 1 is a plan view and a side view of a single support (microparticle) comprising a sequential identification;
  • Figure 2 is a schematic sectional side view of a single support with analytes attached thereto;
  • Figure 4a, 4b are schematic top views diagrams showing the experiment reaction between a substrate with attached primary analytes and secondary analytes bound to supports with identification means before and after wash;
  • Figure 5 is a schematic diagram illustrating a planar-based reader for interrogating the system.
  • Figures 6a, 6b are schematic top views of a planar substrate illustrating examples of the measuring path taken by the planar-based reader of Figure 5.
  • FIG. 1 there is shown an illustration of a preferred embodiment of a support 1 for use in a system according to the invention where the supports 1 are placed on a substrate 10 to allow multiplexed detection of binding activity of analytes attached thereto.
  • a single support 1 such a support 1 will also be referred to as a microparticle or "bead" in the following description.
  • the support 1 can be fabricated f om virtually any insoluble or solid material, for example one or more of polymers, silicates, glasses, fibres, metals or metal alloys, fn the preferred embodiment of the invention, the support 1 is fabricated from a metal, such as gold, silver, copper, nickel, zinc or most preferably aluminium.
  • An associated sequential identification code is thereby recorded in the support 1 as a series of holes using coding schemes similar to those found on conventional bar code systems, for example as employed for labelling merchandise in commercial retailing outlets.
  • Such a code allows the use of existing reader technology to determine the identification 2 of the supports 1, thereby decreasing the initial investment when adopting technology according to the invention.
  • the largest dimension of the supports 1 is smaller than the largest dimension of the area spotted with primary analyte 17 on the substrate 10.
  • the rniniaturisation of the supports 1 decreases the use of reagents and secondary analyte 16 in experiments.
  • With the support 1 smaller than the size of the spot of analyte on the microarray there is also a decreased risk in the supports 1 being associated with more than one spot of analyte when unbound supports 1 have been washed away.
  • the support 1 has suitably a width 4 to length 3 ratio in a range of circa 1:2 to circa 1:20, although a ratio range of circa 1:5 to circa 1:15 is especially preferred. This allows good flow properties of the supports 1 during liquid handling and dispensing.
  • the support 1 has a thickness 5 which is preferably less than circa 3 ⁇ m, and more preferably less than circa 1 ⁇ m.
  • the thickness of less than circa 1 ⁇ m has been shown to provide sufficient mechanical support strength for rendering the support 1 useable in harsh experimental conditions.
  • a preferred embodiment of the invention concerns supports 1 having a length 3 of circa 100 ⁇ m, a width 4 of circa 10 ⁇ m and a thickness 5 of circa 1 ⁇ m; such supports are capable of storing more than 100,000 different identification sequence bar codes 2.
  • Experimental demonstrations of up to 32 bit binary coding schemes theoretically allowing several billion different variants of the supports' 1 codes for use in bioassays for analyte characterization experiments have been undertaken.
  • the identification means 2 is preferably a barcode made up of sequentially arranged holes, which include error checking and directional orientation features to prevent incorrect identification of the support 1. Also end markers 8 of the support's 1 coding system may be used to indicate the intactness or reading direction of the support 1. Supports 1 of different maximum lengths 3 in a range of circa 40 ⁇ m to 100 ⁇ m, carrying between two and five decimal digits of data in the sequential identification 2, have been fabricated for use in different experiments for the detection of analyte characteristics.
  • the shape of the support 1 is such that it optimises the number of supports 1 manufactured per wafer and also substantially optimises the number of identification codes possible on the supports 1.
  • Conventional photolithography and dry etching processes are examples of such manufacturing techniques used to manufacture and pattern a material layer to yield separate solid supports 1 with bar-coded identification 2.
  • the photolithography also allows all types of planar shapes of supports to be manufactured in shapes such as elliptical, circular, rectangular, square, multi-angled and many more.
  • step (5) to facilitate the attachment of an analyte, such as a test sample and or a probe used in multiparameter experimental analysis, to the support 1.
  • the treatment of the supports 1 can be performed after the release from the wafer as described above or alternatively prior to the release from the wafers or earlier in the manufacturing process steps.
  • the treatment of the support material layer, step (5) could be omitted.
  • FIG. 2 shows how secondary analytes 16 are attached to a section 7 of the support 1.
  • the secondary analytes 16 are referred to as test samples, but it will be apparent to the person skilled in the art that the test samples may instead be attached to the substrate 10 with the primary analytes (probes/query molecules) 17 being attached to the supports 1 depending on how experiments are designed and customised.
  • different types of secondary analytes 16 may be attached to supports 1 fabricated by steps (1) to (5) above either before or after executing photolithographic operations or releasing the supports 1 from the planar wafer.
  • the support's 1 surface 6 may also be heated with a polymer material such as silane and/or biotin, to further enhance attachment properties.
  • the supports 1 preferably have silane baked onto their surfaces 6. Attaching linking molecules, for example avidin-biotin sandwich system, to the analytes 16, 17 further enhances their chemical molecular attachment properties.
  • the enhanced attachment is preferably achieved through having electrostatic or more preferably covalent bonds between attachment surface 6 of the support 1 and the secondary analytes 16, so called fixedly attaching the analytes 16 to the supports 1.
  • the covalent bonds prevent the analytes 16 from being dislodged from the supports 1 and causing disturbing background noise during analysis. There is also a potential problem that loose analytes 16 could prevent the identification of reactions that have occurred. It is found to be important to wash the active supports 1, said supports 1 having analytes 16 attached thereto, after attachment to remove any excess analytes 16 that could otherwise increase the noise in the experiment during analysis. Discrimination of the tests is thereby enhanced through a better signal-to-noise ratio.
  • FIG. 3 there is shown schematically a system indicated generally by 9 comprising the substrate (array) 10 and a quantity of liquid 11 including supports 1.
  • the substrate 10 which herein after also is referred to as an array or microarray, has two substantially planar main surfaces 12 and can be of any desired shape, but is most preferably square or rectangular.
  • the substrate 10 may also be made of a variety of materials, such as glass, metal, plastics materials, wafers, membranes or any other material contemporarily used for fabricating microarrays. Most preferably, the substrate 10 is fabricated from a material, for example glass (microscope slide) or plastics material (for example an acrylate), which is light transmissive.
  • the substrate's 10 top main surface 12 is planar or may be divided into sections by partitioning features, for example wells or boundaries, to prevent cross contamination between sections.
  • the main surface 12 of the substrate 10 has preferably a surface area in a range of 0.25 cm 2 to 50 cm 2 , more preferably in a range of circa 1 cm 2 to 25 cm 2 and most preferably in a range of circa 2 cm 2 to 6 cm 2 .
  • On the planar main surface 12 of the substrate 10 primary analytes 17 are fixedly attached by spotting or synthesis.
  • the primary analytes 17 may also be electrostatically or more preferably covalently attached to the substrate 10 as discussed for the supports 1.
  • the primary analytes 17 may then be identified through their position on the substrate 10.
  • the liquid 11, which is placed on the substrate 10, is appropriately a liquid solution and is normally an aqueous solution.
  • the system 9 can be considered to be an assay comprising the liquid solution 11 with secondary analytes 16 loaded supports 1 placed on a substrate 10 with position loaded primary analytes 17.
  • the system 9 is of considerable advantage in that it is capable of providing the benefits of using two dimensional substrates 10 with established reader technology, multiplexing as well as the advantages of the contemporary assays with higher throughput, and good sensitivity and reaction kinetics.
  • analytes 16, 17 When performing a multiparameter analysis of analytes 16, 17 experiments many different types of analytes 16, 17 may be used.
  • the analytes 16, 17 may be antibodies, antigens, proteins, enzyme substrate, carbohydrates, peptides, nucleic acids, peptide nucleic acids, cell lines, chemical components, oligonucleotides, serum components, drugs or any derivatives or fragments thereof.
  • the analytes can be, for example, dyes, preservatives, labelling chemicals (for example for tracking the movement of counterfeit products), radioactive labelling chemicals, and food.
  • Figure 4a shows the analyte loaded substrate 10 with analyte loaded supports 1 before reading and figure 4b shows the same when non reacting supports 1 have been washed away ready for reading.
  • the assay reactions which takes place on the substrate 10 consists of a liquid solution with suspended supports 1 and analytes 16, 17.
  • the analytes are made up of test samples (secondary analytes) 16, probes (primary analytes) 17, and signal emitting labels 18.
  • Many different test samples 16 are used for functioning as reaction molecules in the experiment to be performed, with each type of test samples 16 being attached to a support 1 with a specific identification 2.
  • probe 17 or test sample 16 are bound to each other through a hydrogen bond it allows the bond to be broken separating the substrate with a primary analyte from the support with the secondary analyte, which allows the reuse of the substrate 10.
  • the probes 17 and test samples 16 can be interchanged and used either as primary or secondary analytes depending on the required experiment.
  • the supports' 1 may be the same size as the area of spotted probes 17 on the substrate 10. It is however more preferable for the support 1 to cover less than circa 75%, preferably less than circa 50%, more preferably less than circa 25% and most preferably less than circa 10% of each probe's 17 spotted area on the substrate 10. By the supports 1 being much smaller than the area of spotted probe 17 on the substrate 10, several supports 1 may bind to the spot at any one time.
  • the signal emitting label (reporter system) 18 which has bound to the probes 17 and test samples 16 will then be emitting its signal e.g. fluorescent signal which gives the quantitative measurement.
  • the advantage of using the test sample 16 loaded supports 1 to indicate reaction is that it elhninates the need for several types of reporters.
  • the differently coded supports 1 can be used to indicate the interaction between different probes 17 and test samples 16 while a single fluorescent reporter is used to indicate the quantitative level of the interaction.
  • fluorescent reporter systems to detect multiple interactions on a microarray it becomes difficult to perform multiplexing experiments of many probes 17 against more than 2 different types of test samples 16 as the different fluorescent reporter labels 18 used have problems with spectral overlap and poor results.
  • An example of this embodiment could be the use of several supports 1 with appropriately attached test samples 16 suspended in a liquid solution 11 placed in a well of a 96-well or 384-well ELISA plate 10 with specific probes 17 attached to the bottom of each well. This dramatically improves the throughput of contemporary methods of batching ELISA plates, which pause until a sufficient number of plates are ready for analysis, while still allowing the use of conventional ELISA plate readers.
  • Another example of this embodiment related to the use of a pre-spotted microarray as the substrate 10, while drastically increasing the number of probes 17 analysed through the use of several supports 1 with identification means 2 coated in test samples 16.
  • a further example of this embodiment relates to a microscope slide with multiple probes 17 pre-deposited thereon like a "home brew microarray". This embodiment increases the number of parameters that can be analysed when adding the liquid solution 11 with suspended test sample 16 loaded supports 1 to the probe 17 spotted slide 10.
  • specific secondary analytes 16 are attached to individual supports 1 preferably through covalent bonds. These secondary analytes 16 can, for example, be test samples related to individuals in clinical trials or other research. Multiple secondary analytes 16 on uniquely coded supports 1 can be tested against specific primary analytes (probes) 17 by placing the liquid solution 11 with the suspended supports 1 on a substrate 10 with primary analytes 17 attached thereto. The results of the reaction between probes 17 and test samples 16 will be based on the final position of the supports 1 together with their identification code 2. Using the supports '1 as the only reporter system eliminates the use of any currently reporter system, such as radioactivity or preferably fluorescence to indicate that reaction has occurred.
  • a system 20, illustrated in Figure 5, provides further benefits for all the embodiments of the invention as it allows tailoring of experiments as the supports 1 with test samples 16 may easily be added to the substrate 10 with probes 17 eliminating the long turnaround time needed if using many different substrates 10 for testing against each test sample 16.
  • the system 20 gives an improved flexibility and customisability over conventional microarrays, and increases the throughput allowing multiparameter analysis.
  • the supports 1 are smaller than the deposited primary analyte on the subsfrate 10 to avoid unambiguous analysis and reading. It is also possible to have the test sample as a primary analyte and the probe as a secondary analyte in this embodiment of the invention.
  • a system 9 with primary analytes 17 attached to a substrate 10, such as a slide, microarray or homebrew.
  • Secondary analytes 16 attached to the identifiable supports 1 and suspended in a liquid solution placed on the substrate 10 as in the first and second embodiments above.
  • a tertiary analyte By adding a tertiary analyte to the liquid solution another parameter of analysis can be added to the system. This can be useful if it is difficult to attach a certain analyte to the substrate 10 or support 1.
  • the tertiary analyte may then interact with either the primary or secondary analytes 17, 16, which interaction is shown through the use of a signal emitting label 18, such as a change in colour or fluorescence.
  • the tertiary analyte will have very good sensitivity and reaction kinetics with the secondary analyte 16 as there will be a 3 dimensional interaction between the analytes as the support 1 is suspended in the liquid solution 11.
  • the system then uses the advantages of the existing technologies of 2 dimensional microarrays and 3 dimensional solution based arrays.
  • the different analytes may be e.g. either probes 17 or test samples 16 as outlined above.
  • This embodiment of the invention is comparable to a competitive ELISA experiment with quantitative measurements where there is competition between the different analytes when interacting.
  • a reading means used for reading the subsfrate 10 with loaded supports 1 suspended thereon in a liquid solution 11 will now be described with reference to Figures 5, 6a and 6b.
  • the liquid solution 11 with secondary analyte 16 loaded supports 1 have been placed on the primary analyte 17 loaded substrate 10
  • movement of the supports 1 over the substrate's 10 main surface can be done using established laboratory equipment such as rockers, agitators, shakers or belly dancer. This is to assure that the secondary analytes 16 of the supports 1 are given the opportunity to interact with all the primary analytes 17 on the substrate 10.
  • the reaction time has been sufficient allowing full interaction between the analytes 16, 17 the substrate 10 is washed removing all the secondary analyte bound supports 1 and signal indicators 18 before the substrate 10 is read and analysed.
  • the test result of reacting analytes 16, 17 is measured as a yes/no binary result or by the degree of fluorescence 10 emitted from the signal emitting label 18.
  • the system 20 consists of a reader, as shown in figure 5.
  • the reader includes a measuring unit indicated by 25 which measures the identification 2 of the supports 1 tagged to analytes 16.
  • the measuring unit has a detection unit 27 to detect the fluorescent reaction signal 19 form the interacted analytes 16 and a reader unit 30 to read the identification code 2 of the supports 1.
  • the detection unit 27 has a fluorescence microscope when detecting the fluorescent signal 19 indicating reaction.
  • the reader unit 30 has a barcode reader to read the transmissive barcodes 2 of the supports 1. If required to have multiple reporter systems it is preferable to have different type of signal for the support 1 identification 2 and the reaction detection 19, as there then is a limited risk of the signals being mixed up or being overlapping (spectral overlap). This allows for greater multiplexing (multiple simultaneous reactions) possibilities.
  • the best solution for qualitative analysis is to use the support 1 with identification 2 as the only reporter system eliminating all the need for any radioactive or fluorescent reporters needing mercury lamps or similar to be able to detect the reporter signal.
  • the support 1 may also be detected on the subsfrate 10 in reflective or transmissive light using a bright field light source.
  • a processing unit 28 of the measuring unit 25 calculates the results of the tests associated with the supports 1. This sufficient number is preferably between 10 and 100 copies of each type of supports 1; this number is preferably to enable statistical analysis to be performed on test results. For example, statistical analysis such as mean calculation and standard deviation calculation can be executed for fluorescence 10 associated with each type of test sample 16 and/or probe 17 present.
  • a processing unit 28 is also included for controlling the detector and reader units 27, 30 so that the each individual support 1 is only analysed once.
  • the software of the processing unit 28 can preferably be configured to analyse only the supports 1 that give off a signal 19, for example through a fluorescent signal label 18, indicating that an interaction hetween the analytes 16, 17 characteristics has occurred. This could also be used to avoid detection of supports 1 with secondary analytes 16 which have not reacted with the primary analytes 17 on the substrate 10 but not been washed off prior to analysis.
  • the analysis of the loaded substrate 10 using the measuring unit 25 is a very cost effective, easy to perform and suitable way to multiply the analysing capacity for low to medium sample numbers in the range of, for example, single figures to a few thousand supports 1 on each substrate 10.
  • Figure 6a and 6b Preferred paths 50 for systematically interrogating the substrate 10 are shown in Figure 6a and 6b.
  • Figure 6a is a depiction of a meander-type interrogation regime
  • Figure 6b is a depiction of a spiral-type interrogation regime.
  • paths 50 apparent to one skilled in the art, for example moving the substrate 10 in an opposite direction to the path 50, moving the subsfrate in a meandering diagonal path, or covering the whole substrate in one substantially linear path across its surface.
  • the regimes of Figures 6a, 6b are efficient for achieving an enhanced support 1 read speed.
  • a stepper-motor actuated base plate 40 supporting and bearing the subsfrate 10 may be used to move the substrate 10 around while the measuring unit 25 is held stationary.
  • the measuring unit's 25 reader unit 30 for image-processing is used to capture digital images of each field of the subsfrate 10 with a liquid solution 11 suspending supports 1 with attached analytes 16 thereon. Digital images thereby obtained correspond to light transmitted through the substrate 10 and past a base plate 40 and then through the supports 1 rendering the supports 1 in silhouette view; such silhouette images of the supports 1 are analysed by the reader unit 30 in combination with a processing unit 28.
  • the sequential identification 2, for example a bar-code, associated with each support 1 is hence identified from its transmitted light profile by the reader unit 30.
  • the signal emitting unit 29 generates a fluorescent signal, which signal makes the labels 18 on supports 1 fluoresce indicating a positive reaction 19 between a test sample 16 and a probe 17.
  • a detector unit 27 detects the magnitude of fluorescence 19 from activated supports 1 to identify the degree of reaction.
  • the fluorescent signal 19 integrated over activated supports' 1 surface 6 is recorded in association with the identification bar-code
  • the processing unit 28 is connected to the light source 45, the signal unit 29, the reader unit 30, and the detector unit 27 and to a display 46. Moreover, the processing unit 28 comprises a control system for controlling the light source 45 and the signal unit 29.
  • the light silhouette and fluorescent signals 19 from the supports 1 pass via an optical assembly 41, for example an assembly comprising one or more lenses and or one or more mirrors, towards the detector unit 27 and reader unit 30.
  • a mirror 42 is used to divide the optical signals into two paths and optical filters 43, 44 are used to filter out unwanted optical signals based on their wavelength.
  • the light source 45 and signal unit 29 can be turned on and off at intervals, for example mutually alternately. Signals are received from the reader unit 30 and detector unit 27, which are processed and corresponding statistical analysis results presented on a display 46.
  • the intended uses of the system 20 may be in any process where experiments requiring the analysis of multiparameter analysis of analytes.
  • the applications where several parameters are involved are for example in biochemical detection of one or more analyte characteristics including gene expression, SNPs analysis, nucleic acid testing, antibody or protein analysis, lead target identification and drug targeting.
  • biochemical detection of one or more analyte characteristics including gene expression, SNPs analysis, nucleic acid testing, antibody or protein analysis, lead target identification and drug targeting.
  • the primary analytes 17 and secondary analytes 16 which can be either test samples or probes can be interchanged so that there is a multiparameter analysis experiment possible irrespectively which analyte is attached to the coded support 1 or the position based coded substrate 10.
  • the size of the supports 1 should be less than the half of the smallest distance between to spots of deposited analyte on the substrate 10 to avoid overlapping supports 1 attached to different spots.

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Abstract

L'invention concerne un système d'analyse multiparamétrique de substances à analyser, comprenant les éléments suivants: 1) un substrat solide (10) dont la surface principale (12) s'étend sensiblement dans un plan bidirectionnel; 2) au moins un analyte primaire (17) lié à la surface principale (12) du substrat (10), la surface principale (12) pouvant recevoir un liquide (11) contenant au moins un analyte secondaire (16); et 3) des appareils de mesure (27, 29, 30) disposés en contact visuel avec la surface principale (12) du substrat solide (10) pour surveiller cette surface (12). Le système est caractérisé en ce que: 4) au moins un support (1) est suspendu dans le liquide (11); 5) le support (1) comporte une identification (2) séquentielle pour l'identifier; 6) au moins un analyte secondaire (16) est attaché au support (1); et 7) les appareils de mesure (27, 29, 30) sont placés pour détecter toute interaction post-réactionnelle entre un ou plusieurs analytes primaires (17) et un ou plusieurs analytes secondaires (16) sur la base de l'identification et de la position finale du support (1) sur le substrat (10). La présente invention porte également sur un procédé d'analyse multiparamétrique de substances à analyser au moyen de ce système.
PCT/GB2003/001725 2002-04-24 2003-04-23 Systeme et procede d'analyse multiparametrique de substances a analyser WO2003091731A1 (fr)

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AU2003224297A AU2003224297A1 (en) 2002-04-24 2003-04-23 System and method for multiparameter analysis of analytes

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GB0209319A GB2387903A (en) 2002-04-24 2002-04-24 Multiparameter analysis using tagged molecules
GB0209319.3 2002-04-24

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Cited By (5)

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