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WO2008122241A1 - Analyses rapides de protéines et dispositif associé - Google Patents

Analyses rapides de protéines et dispositif associé Download PDF

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Publication number
WO2008122241A1
WO2008122241A1 PCT/CN2008/070671 CN2008070671W WO2008122241A1 WO 2008122241 A1 WO2008122241 A1 WO 2008122241A1 CN 2008070671 W CN2008070671 W CN 2008070671W WO 2008122241 A1 WO2008122241 A1 WO 2008122241A1
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WIPO (PCT)
Prior art keywords
membrane
molecules
target
lateral flow
detection
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PCT/CN2008/070671
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English (en)
Inventor
Joseph Wing On Tam
Wai Hon Chan
Joseph K. F. Chow
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Diagcor Bioscience Inc Ltd
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Diagcor Bioscience Inc Ltd
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • 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
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0877—Flow chambers

Definitions

  • the present invention relates to methods and devices for making rapid, definitive identification of different protein profiles by arrays analyses using a flow-through process.
  • the typing classification can be set from low (degenerate) to complete differentiation.
  • standard serological typing Keratige et . al . , 1993; Chow and Tonai, 2003; Mach et al . , 2004
  • dot-blot, reverse dot-blot or slot blot can be used for other protein systems for rapid analysis described in this invention.
  • the present invention provides a lateral flow-through device for protein analysis, comprising: (a) one or more reaction chambers, each of which comprises a membrane for immobilizing capture molecules capable of capturing target analysts; and (b) liquid delivery elements capable of accepting and removing solution comprising the target analysts to and from the reaction chambers, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising the capture molecule to the width of the membrane.
  • the present invention provides a lateral flow-through device for protein analysis, comprising: (a) one or more membrane cassette assemblies, each of which comprises a plurality of wells, wherein each of the wells comprise a membrane for immobilizing capture molecules capable of capturing target analysts; and (b) liquid delivery elements capable of accepting and removing solution comprising the target analysts to and from the membrane cassette assemblies, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising the capture molecule to the width of the membrane .
  • the devices described above further comprise (a) controlling elements that can be regulated to maintain the reaction chamber or membrane cassette assembly in controlled conditions; and (b) connecting elements for connection to a power supply and control unit that can regulate and maintain the controlled conditions.
  • the present invention also provides a lateral flow- through protein analysis system comprising a pluraity of the devices described above, wherein the devices are connected to a power supply and control unit capable of supplying energy and providing regulatory control to the devices.
  • the present invention also provides a method of performing rapid protein detection, comprising the steps of: (a) immobilizing capture molecules on membranes placed in any one of the devices described above, wherein the capture molecules are capable of capturing one or more target protein molecules on the membranes; (b) applying to the membranes a sample comprising the target protein molecules, wherein the sample is maintained in a lateral flow direction that allows the sample flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising a capture molecule to the width of the membrane; and (c) detecting the captured target protein molecules on the membranes .
  • Figure 1 shows an exploded view of a lateral flow- through device of the present invention:
  • Figure IA shows a device containing a reaction chamber where the membrane is placed directly onto the thermal plate which consists of the heating unit and the sensor regulator and electrical leads to the central power unit [P] as indicated in the Figure IB.
  • Figure IB shows an arrangement of multiple lateral flow-through detection devices (I), (II) and (III) etc. which can be regulated when connected to a central control unit [P] .
  • the sample and reagent solutions shall be delivered from the top cover openings and flow across the membrane for target molecules capture and development or additional assembly for solution removal i.e. at the drainage outlets .
  • Figure 1C shows a detailed view of another embodiment of a flow-through reaction chamber assembly. This embodiment is designed for use in the direct flow-through device of the prior art.
  • the sample as well as reagent solutions flow through the incubation well [I] along the direction of the arrow ⁇ through the reaction chamber [R] .
  • Reaction solution is drawn out by the suction (drainage system) provided by the main device unit, followed by signal development and interpretation.
  • Figure 2 shows another embodiment of a lateral flow reaction chamber assembly as described herein.
  • Figure 3 shows cancer biomarkers commonly used for clinical diagnosis. Quantitative assays are usually done with ELISA separately for each of these markers. Proteomic markers are identified and determined either by mass spectrometry (MS) and/or 2 dimensional gel electrophoresis as described in the given literatures .
  • MS mass spectrometry
  • 2 dimensional gel electrophoresis as described in the given literatures .
  • Figure 3A shows plasma/serum concentration ranges of cancer biomarkers and their cut-off values among normal individuals reported from various studies.
  • FIG. 4 shows examples of array images of cancer biomarkers.
  • Panels A-D are image profiles obtained from some cancer samples;
  • panel E is data from a normal sample.
  • FIG. 5 shows example of concentration gradient and the detection limit of the present flow-through method for protein array detection.
  • Figure 6 shows a typical protein array profile (relative concentrations of different biomarkers) and biomarkers used for liver cancer diagnosis.
  • Figure 7 shows dot-blot analyses in a lateral flow- through manner of several lysates in identifying the culture conditions that generate positive signals (Figure 7A) as a mean for rapid screening for protein expression studies followed by Western Blot for detailed protein identification ( Figure 7B) .
  • PC 12 cells were stimulated with NGF for 0'; 15'; 30'; lhr; 3hrs; 5hrs and 24 hrs .
  • Lysates were screened by dot blot assays with anti-ERK MAP kinase with the flow-through device as shown in Figure 7A.
  • parts of the same lysates were subjected to 10% PAGE, followed by conventional blotting procedures and flow-through procedures.
  • Figure 7B shows the Western Blot analyses of lysates for the corresponding culture conditions. Evidently the results from rapid screening correlate well with results obtained by Western in positive signal for final identification of the protein (s) induced in given conditions. The dot-blot screening time was 30-40 minutes whereas the total Western Blotting assay required a two-days process that include protein separation and transfer etc. Figure 7B also showed that the results obtained by conventional process (which requires over two hours) compared well with the results obtained from flow- through process in 30 minutes at much reduced reagents used.
  • a lateral flow-through device for protein analysis comprising: (a) one or more reaction chambers, each of which comprises a membrane for immobilizing capture molecules capable of capturing target analysts; and (b) liquid delivery elements capable of accepting and removing solution comprising the target analysts to and from the reaction chambers, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising the capture molecule to the width of the membrane.
  • the reaction chambers are disposable.
  • the present invention provides a lateral flow-through device for protein analysis, comprising: (a) one or more membrane cassette assemblies, each of which comprises a plurality of wells, wherein each of the wells comprise a membrane for immobilizing capture molecules capable of capturing target analysts; and (b) liquid delivery elements capable of accepting and removing solution comprising the target analysts to and from the membrane cassette assemblies, wherein the solution is maintained in a lateral flow direction that allows the solution flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising the capture molecule to the width of the membrane .
  • liquid delivery elements capable of accepting and removing solution comprising the target analysts to and from the reaction chambers or membrane cassette assemblies.
  • the solution is drawn across the membrane by absorbance wicks on one side.
  • accepting and removing solution to and from the reaction chamber or membrane cassette assembly can be done by regulated liquid pumping.
  • the liquid pumping is used to recirculate the solution comprising the target analysts through the membrane.
  • the devices described above further comprise (a) controlling elements that can be regulated to maintain the reaction chamber or membrane cassette assembly in controlled conditions; and (b) connecting elements for connection to a power supply and control unit that can regulate and maintain the controlled conditions.
  • the power supply and control unit is capable of supplying energy and providing regulatory control to maintain the reaction chambers in the controlled conditions.
  • the controlling elements are heating or cooling elements.
  • the membrane can be made of nitrocellulose, nylon, Nytron, Biodyne, Porex or any kind of porous matrix material support, and the capture molecules can be arranged in an array on the membrane .
  • the present invention also provides a lateral flow- through protein analysis system comprising more than one of the devices described above, wherein the devices are connected to a power supply and control unit capable of supplying energy and providing regulatory control to the devices.
  • a power supply and control unit capable of supplying energy and providing regulatory control to the devices.
  • each of the devices is controlled independently by the power supply and control unit so that each device can perform different analysis under different conditions.
  • the present invention also provides a method of performing rapid protein detection, comprising the steps of: (a) immobilizing capture molecules on membranes placed in any one of the devices described above, wherein the capture molecules are capable of capturing one or more target protein molecules on the membranes; (b) applying to the membranes a sample comprising the target protein molecules, wherein the sample is maintained in a lateral flow direction that allows the sample flows through the membranes in cross-section so that the sensitivity of detection is dependent on the ratio of the width of an area comprising a capture molecule to the width of the membrane; and (c) detecting the captured target protein molecules on the membranes.
  • the target protein molecules are of human, bacterial, or viral origin.
  • the human is having or is suspected of having cancer.
  • the captured target protein molecules are detected by a method such as fluorescence tags, quantum dot labeling, colloidal gold particle labeling, magnetic particle labeling, or enzyme-linked substrate assay.
  • the target protein molecules are mixed with a signal generating agent before being applied to the membranes for capture detection.
  • the target protein molecules are biomarkers for cancer, therefore detection of the target protein molecules would provide diagnostic information for cancer.
  • the method described herein can be used to provide diagnostic information for liver cancer when the capture molecules are capable of binding to target protein molecules comprising AFP, AFP-L3, CA19.9, CA125, CA15.3 , TGF-Bl, HS-GGT, ALF-161, des-gamma carboxyprothrombin, ferritin, HbsAg, and anti-HCV antibody.
  • diagnosis or prognosis of liver cancer is indicated by an increase in expression levels as compared to normal cut-off values for two or more target proteins selected from the group consisting of AFP, AFP-L3, HS-GGT, TGF-Bl and DCP.
  • the increased expression levels are detected in a sample obtained from a human infected with hepatitis B virus, hepatitis C virus, or hepatitis B and C viruses.
  • the method described herein can be used to provide diagnostic information for a specific cancer when the capture molecules are capable of binding to two or more of the biomarkers as listed in Figure 3.
  • diagnosis or prognosis of cancer is indicated by an increase in expression levels as compared to normal cut-off values for two or more of the biomarkers as listed in Figure 3.
  • the present invention also provides a method of concentrating target molecules from large volume and detect low concentration of target molecules, comprising the steps: (a) the sample is first incubated with secondary affinity molecules such as substrates for enzymes; ligands for receptors; secondary antibodies for antibodies etc. immobilized solid matrix; (b) separate matrix and remove sample solution; (c) re-dissolve target molecule from matrix follow by (d) flow-through detection with reduction of non-specific binding for increase in signal to noise for increased sensitivity.
  • secondary affinity molecules such as substrates for enzymes; ligands for receptors; secondary antibodies for antibodies etc.
  • the present invention also provide a method for rapid and high throughput screening of protein expression profile followed by Western Blotting identification, comprising (a) antibodies array screening membrane; and (b) applying in the flow- through manner multiple lysates of cell cultures having different conditions of inductions for either up-regulated or down regulated expression in identifying component (s) in regulatory factor for biochemical pathways; (c) detecting and identifying changes of expression and identifying the culture conditions responsible for the changes which enables the investigator to have tremendous increase in speed and efficiency to explore the complex biochemical world and (d) follow the conventional time consuming Western Blotting experiments for molecular identification only a limited fraction of experiments.
  • the method provided by this invention would be of utmost importance for a saving of many folds in time and material cost for research.
  • Figure 1 shows an exploded view of a lateral flow-through detection device of the present invention, and possible arrangement of multiple lateral flow-through detection devices connected to a central control unit.
  • the device comprises a central controlling unit connected to one or more lateral flow-through devices.
  • the central controlling unit provides power to and controls the lateral flow device where protein binding reactions and developing procedures are carried out.
  • Several reactions or several samples and/or analytes) can be tested simultaneously in a single lateral flow device, or in several devices controlled individually at different conditions.
  • the lateral flow device can accommodate an array in a format of n x m dot matrix (array) or in the form of linear arrays (as shown in Figure 1) .
  • array n x m dot matrix
  • Figure 1 linear arrays
  • a test solution flows from one end of the array to the other end of the array (i.e., in an east to west, or in a north to south direction), hence the sensitivity of detection is increased substantially.
  • the extent of increase in sensitivity depends on the ratio of the total area of the array/membrane to the area of the dot or line containing the capturing probes. For example, assuming the total area of an array/membrane is 100mm square, and the dot size is lmm square.
  • the sensitivity is only dependent on the ratio of the width of the dot to the width of the membrane (i.e., the cross section of the membrane) .
  • the total amount of solution that will pass through a 1 mm dot provided on a 10mm x 10mm membrane will be about 1/10, which represents a 10-fold increase in sensitivity using the same amount of test solution containing the target molecules.
  • the sensitivity will also be increased since all the target molecules will pass through the line extending across the strip (or membrane) .
  • the lateral flow-through process allows quantitative measurements to be taken during the reaction process because the flow of the analyst is more uniform.
  • FIG. IA One embodiment for the lateral flow-through device is shown in Figure IA.
  • This embodiment can be made as a new device that comprises (i) an electronic control unit for operations; (ii) an optional temperature block; (iii) a reaction chamber; and (iv) a liquid delivery system.
  • Temperature control may not be essential for antibody-antigen reaction as generally done for Western Blot analyses. However, some proteins may be less stable, others may react better at certain temperature higher or lower than room temperature, therefore accurate controlled conditions including temperature is one embodiment. On the other hand, if less stringent condition is favorable for certain analytes detection, a simpler device is in order for a low cost alternative device .
  • the reaction chamber where the membrane is located can be designed as a new separate and/or disposable unit.
  • the flow direction, the speed of flow and the sequence of solution reagents flow can be controlled accurately by the controller.
  • This disposable reaction chamber can be made as single or multiple cassettes for separate or parallel reactions to provide increase in throughput.
  • this embodiment can provide optimal conditions for effective detection with increase in sensitivity and specificity.
  • the disposable as well as the totally enclosed setting in this embodiment can prevent any possibility of cross contamination.
  • Alternative embodiment for the lateral flow-through device shown in Figure 1C is modified assembly of the reaction chamber which enables one to adopt using the Direct Flow-through Device described in the prior invention [U.S. patent No. 6,020,187] .
  • the assembly comprises a plurality of wells, each of which further comprises a membrane.
  • the cassette like membrane assembly by which the sample and subsequent reagent solutions can be applied on top of one side of the array membrane where the solution will flow across the membrane and drained from the other side of the membrane into the waste. Recirculation is also possible provided re-circulating pumping mechanism is added.
  • the flow-through array system described herein can be use for dot-blot, reversed dot-blot or slot blot analysis, where multiple array assays can be done simultaneously.
  • target samples to be tested are dotted onto the membrane as arrays in each well
  • a membrane refers to any porous matrix materials capable of binding the target antigens for detection.
  • Slot Blots can be done either as Dot-blot or Slot-Blot as described above.
  • a set of biotin labeled at various concentrations can be immobilized onto the membrane. Color development is done according to established procedures with avidin-alkaline phosphatase system. The lowest detection limit of the immobilized antigen is used as the lowest limit of detection for the present embodiment of the flow-through device.
  • the flow-through reaction is determined by immobilizing various antibodies forming the specific array as capture molecules. Various concentrations of present gradient of target molecules are used to test the feasibility and effectiveness of quantitative capturing process by the immobilized antibodies for their corresponding target molecules.
  • the present invention provides a rapid platform (the lateral flow-through device's reaction chamber) for carrying out all the steps of Western Blot after the proteins have been transferred onto a membrane.
  • the lateral flow-through process by itself is suitable for such improvement by reducing the time and reagent cost by several folds simply by performing steps (3) and
  • the membrane is placed onto the reaction chamber of the present lateral flow-through device, and start with the normal conventional procedures: (i) apply blocking solution and allow to flow into the membrane preventing non-specific binding; (ii) add antibody conjugate, add washing solution to remove unbound conjugates; and (iii) add substrate for signal development and analyze data.
  • the platform of the present invention will provide rapid immuno-reaction between target molecules and their reactants
  • the present invention provides an antibody array and the lateral flow-through system as a first tool for qualitative screening analyses. Detailed analyses will be done only on those lysates that are identified to have significant changes. For illustration purpose, the MAP kinase activation is used as an example.
  • Antibody array was prepared by immobilizing individual antibodies onto the membrane. The time required for performing 15 wells reactions (i.e.
  • the present lateral flow-through system can be used for rapid initial screening for the presence or absence of target proteins before performing the long electrophoresis separation and Western analyses.
  • Using the new lateral flow-through system and dot -blot to screen many samples for target protein requires only 20-30 minutes without performing conventional electrophoresis separation, transfer and Western Blot analysis.
  • the flow-through system of the present invention will save 10-100 folds in time and materials. Furthermore, using a recirculation flow-through system disclosed herein would even further increase the sensitivity of the assay.
  • the Flow-through Array described herein can generate useful profiles by simultaneously assaying multiple biomarkers in a single assay.
  • Figure 3 shows some of the biomarkers useful for cancer screening in clinical laboratories for the past decade or two as single marker assays. Although an individual marker may have some prognostic value, a single marker alone cannot provide significant sensitivity and specificity for the diagnosis of solid cancers. Quantitative profiles of a group of markers would be much more useful in delineating possible types of cancer and to get a more accurate early diagnosis.
  • Figure 4 shows a typical set of profiles and their corresponding cancers suggest that such an approach is feasible.
  • BIOMARKERS C-12 cancer screening kit has claimed an advantage of increased detection. Since most of those markers are non-specific for cancer, increase in detection may also be caused by non-cancerous diseases. For example, most liver cancer patients have abnormally high blood serum levels of alpha- fetal protein (AFP) , yet similar increase can also be caused by cirrhosis or chronic hepatitis. Hence further increase in sensitivity of detection will also lead to increase in false positive predictions. Indeed recent general screening data using the C-12 screening test have resulted in over 30% positive rate, which is 10 times the cancer prevalence rate of 3% from the China national statistic data, indicating that the false positive rate is much too high.
  • AFP alpha- fetal protein
  • microarrays capable of analyzing thousands of genes differentially expressed between cancerous and normal patients are used to identify biomarkers responsible for the set cancer as diagnostic tool (MacBeath G 2000; Nguyen et al . , USPTO No. 7049151; Diamond; Scott L, USPTO No. 7332286). Liew CC. et al .
  • biomarkers which include amyloid beta precursor-like protein 2(APLP2); BCL2-related protein Al (BCL2A1) ; phosphoprotein regulated by mitogenic pathways (C8FW) ; complement Component 5 (C5) ; CD14 antigen (CD14) and many others including RAS viral gene sequences and expressed said markers for use as diagnostic tool for liver cancer.
  • APLP2 amyloid beta precursor-like protein 2
  • BCL2A1 BCL2-related protein Al
  • C8FW phosphoprotein regulated by mitogenic pathways
  • C5 complement Component 5
  • CD14 antigen CD14
  • the flow-through device of the present invention is an efficient tool for performing the protein arrays analysis which would provide more sensitive and specific results.
  • the array profiles and diagnosis can be done and further validated through large clinical trails.
  • the present invention can simultaneously detect multiple proteins of different organisms such as viruses and/or bacteria. For example, phenotyping of drug resistant proteins in human such as P450; mutant oncogenic proteins such as Ras, p53 mutated proteins or viral proteins of HIV, HBV, HCV can be conducted singularly or in combinations .
  • liver cancer is one of the most prevalent cancer in China and Africa. Because it is difficult to detect at early stages, once diagnosed the 5-year survival rate is only 5-7%. Hence it is imperative to find more sensitive and specific assays for early diagnosis.
  • serum levels of alpha- fetal protein (AFP) are most frequently used as the prognostic marker because between 50-75% of HCC have abnormally high serum levels of AFP.
  • AFP alpha- fetal protein
  • liver specific HS-AFP or AFP-L3 Li et al . , 2001; Wu W. et al., 2006
  • Midkine USPTO No.
  • the present invention provides in this example a flow-through array comprising a set of biomarkers associated with tumorgenic release from the cancerous cells and this set of markers together with known cell specific mutagenic agents form an array that can serve as diagnostic assay for liver and other cancers.
  • the markers used for liver cancer diagnosis include AFP; AFP-L3; CA19.9; CA125; CA15.3; TGF- Bl; HS-GGT; ALF-161; DCP (des-gamma carboxyprothrombin) ; ferritin; HbsAg; and anti-HCV antibody.

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Abstract

La présente invention concerne des procédés d'identification rapide et définitive de différents profils de protéines par des analyses de jeux utilisant un processus sur membrane en écoulement continu et les dispositifs associés. L'invention concerne notamment des jeux de transfert dot-blot, slot-blot ou dot-blot inverse permettant d'obtenir un criblage à haut débit. L'invention concerne également des procédés et des dispositifs comprenant des ensembles de biomarqueurs pour le diagnostic de cancers.
PCT/CN2008/070671 2007-04-04 2008-04-03 Analyses rapides de protéines et dispositif associé Ceased WO2008122241A1 (fr)

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Cited By (13)

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US7732138B2 (en) 2001-11-07 2010-06-08 Diagcor Bioscience Incorporation Limited Rapid genotyping analysis and the device thereof
CN102879567A (zh) * 2012-09-29 2013-01-16 同昕生物技术（北京）有限公司 用于肝癌诊断的甲胎蛋白异质体分离试剂盒及其组成试剂与应用
CN103293295A (zh) * 2013-05-07 2013-09-11 上海爱纳玛斯医药科技有限公司 检测hbv的磁性生物探针、试纸条及其制备和使用方法
CN103558381A (zh) * 2013-11-06 2014-02-05 昆明云大生物技术有限公司 检测人类艾滋病病毒抗体的免疫层析试纸及其制备方法
CN103558399A (zh) * 2013-11-06 2014-02-05 昆明云大生物技术有限公司 检测乙肝表面抗原的免疫层析试纸及其制备方法
CN103822878A (zh) * 2012-11-16 2014-05-28 上海市肿瘤研究所 凝集素功能化的纳米金及其制备方法和应用
US8980555B2 (en) 2010-04-29 2015-03-17 Diagcor Bioscience Incorporation Limited Rapid genotyping analysis and devices thereof
CN104569401A (zh) * 2014-12-10 2015-04-29 浙江工业大学 一种ca15-3检测试剂盒及其应用
CN104991068A (zh) * 2015-07-02 2015-10-21 天津大学 一种基于量子点的甲胎蛋白免疫层析试纸条的制备方法
CN104991063A (zh) * 2015-06-25 2015-10-21 天津大学 基于量子点的癌胚抗原免疫层析试纸条的制备方法
CN105044340A (zh) * 2015-07-02 2015-11-11 天津大学 一种基于量子点的前列腺特异抗原免疫层析试纸条的制备方法
CN105445470A (zh) * 2014-09-01 2016-03-30 江苏泽成生物技术有限公司 糖类抗原ca125定量测定试剂盒及其制备方法与检测方法
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CN104569401A (zh) * 2014-12-10 2015-04-29 浙江工业大学 一种ca15-3检测试剂盒及其应用
CN104991063A (zh) * 2015-06-25 2015-10-21 天津大学 基于量子点的癌胚抗原免疫层析试纸条的制备方法
CN104991068A (zh) * 2015-07-02 2015-10-21 天津大学 一种基于量子点的甲胎蛋白免疫层析试纸条的制备方法
CN105044340A (zh) * 2015-07-02 2015-11-11 天津大学 一种基于量子点的前列腺特异抗原免疫层析试纸条的制备方法
WO2020224344A1 (fr) * 2019-05-05 2020-11-12 南通大学附属医院 Dispositif et procédé de détection d'immunoempreinte dot
US12461104B2 (en) 2019-05-05 2025-11-04 Affiliated Hospital Of Nantong University Detection device and detection method of dot immunoblotting detection

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