WO2014153262A1 - Filets moléculaires sur phases solides - Google Patents

Filets moléculaires sur phases solides Download PDF

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Publication number
WO2014153262A1
WO2014153262A1 PCT/US2014/029823 US2014029823W WO2014153262A1 WO 2014153262 A1 WO2014153262 A1 WO 2014153262A1 US 2014029823 W US2014029823 W US 2014029823W WO 2014153262 A1 WO2014153262 A1 WO 2014153262A1
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WIPO (PCT)
Prior art keywords
analyte
molecular
molecular net
solid phase
capture
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PCT/US2014/029823
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English (en)
Inventor
Emily Stein
Bruce Phelps
Robert PLACE
Dina UZRI
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Sevident, Inc.
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Publication date
Priority claimed from US13/938,055 external-priority patent/US9910040B2/en
Application filed by Sevident, Inc. filed Critical Sevident, Inc.
Priority to EP14768268.6A priority Critical patent/EP2972343A4/fr
Priority to CA2908613A priority patent/CA2908613A1/fr
Priority to CN201480025377.0A priority patent/CN105339791B/zh
Priority to AU2014236090A priority patent/AU2014236090B2/en
Publication of WO2014153262A1 publication Critical patent/WO2014153262A1/fr

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    • 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
    • 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
    • G01N33/54353—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand attached to the carrier via a chemical coupling agent

Definitions

  • analyte capture technology for solid phase surfaces or devices that can selectively capture analytes from a complex sample with little or no sample preparation and to position said selected analytes in a manner to maximize captured analyte measurement and/or detection in a manner that is compatible with most technologies.
  • a device may comprise a solid phase and a molecular net coupled to at least a portion of a surface of the solid phase.
  • the molecular net may include capture molecules of at least one type coupled to each other by linker molecules of a plurality of types to form a covalently-linked multi-layered three- dimensional matrix.
  • the capture molecules may be configured to bind to the analyte.
  • a method may comprise providing a solid phase, and placing a molecular net on at least a portion of a surface of the solid phase.
  • the molecular net may include capture molecules of at least one type coupled to each other by linker molecules of a plurality of types to form a covalently-linked multi-layered three-dimensional matrix.
  • the capture molecules may be configured to bind to the analyte.
  • a method may comprise providing one or more devices each comprising a solid phase and a molecular net covering at least a portion of a surface of the solid phase.
  • the molecular net may include capture molecules of at least one type coupled to each other by linker molecules of a plurality of types to form a covalently-linked multi-layered three-dimensional matrix.
  • the capture molecules configured to bind to the analyte.
  • the method also comprises exposing the devices to the sample and allowing at least a portion of the analyte to bind to the capture molecules of the molecular nets of the devices.
  • FIG. 1 shows a comparison of traditionally conjugated microparticles
  • FIG. 2 shows a traditional capture molecule conjugation to microparticles and the corresponding analyte measurement capability.
  • FIG. 3 shows an effectiveness of Molecular Net microparticles in measuring analyte.
  • FIG. 4 shows an effectiveness of a Molecular Net with topology in measuring analyte.
  • FIG. 5 shows a comparison of traditionally conjugated microparticles
  • FIG. 6 shows a comparison of traditionally conjugated microparticles
  • FIG. 7 shows an exemplary Molecular Nets on particles.
  • FIG. 8 shows an exemplary Molecular Net topological features on particles.
  • FIG. 9 shows an exemplary Molecular Nets for analyte delivery.
  • Properties of Molecular Nets may be imparted by: the capture molecules selected for use (examples of capture molecules may include antibodies, nucleic acid probes, enzymes, recombinant proteins, peptides and others); the resultant specificity said capture molecules impart; the size and number of selected capture molecules; the placement and spacing of the capture molecules in the molecular Net layer(s); the combination of capture molecules; the order in which the capture molecules may be used; and the ratio of capture molecules to linker molecules and spacer molecules used.
  • the capture molecules selected for use examples of capture molecules may include antibodies, nucleic acid probes, enzymes, recombinant proteins, peptides and others
  • the resultant specificity said capture molecules impart; the size and number of selected capture molecules; the placement and spacing of the capture molecules in the molecular Net layer(s); the combination of capture molecules; the order in which the capture molecules may be used; and the ratio of capture molecules to linker molecules and spacer molecules used.
  • Linker molecules selected for use include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the chemical specificity of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the Angstrom length of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the chemical specificity of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the Angstrom length of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the chemical specificity of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the Angstrom length of the linker molecules include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • the combination of linker molecules include order in which the linker molecules may be used
  • the ratio of capture molecules to linker molecules and spacer molecules used include homobifunctional, heterobifunctional, trifunctional and multifunctional types
  • Properties of Molecular Nets may also be imparted by: the spacer molecules selected for use (examples of spacer molecules include PEG, polymer, nucleic acid, albumin, Fc region, peptide, and other); the chemical properties of the spacer molecules; the size and number of spacer molecules; the order in which the spacer molecules may be used; and the ratio of spacer molecules to linker molecules and capture molecules used.
  • Placement and spacing of the capture molecules, linker molecules and spacer molecules may: confer a characteristic topology on the Molecular Net surface; confer a characteristic density within each layer of a Molecular Net; confer a characteristic porosity of a Molecular Net; remove spatial constraints and thus stearic hindrance; improve binding capacity; reduce non-specific binding; enable the binding of multiple forms of analyte (for example, simultaneous capture of degraded analyte, whole analyte and complexed analyte), and other.
  • the porosity within a Molecular Net may be random, pseudorandom or irregularly interspersed. Porosity of a Molecular Net may be used to filter a sample; may be used to discriminate binding potential molecules in a sample by size-exclusion; may be used to enable macromolecular or cellular binding due to the reduction in stearic hindrance, or other.
  • the pores of a Molecular Net comprise capture molecules, linker molecules and may comprise spacer molecules.
  • Traditional approaches to generate porosity on a solid phase relates to the mechanical modification of the surface of the solid phase and employ methods such as laser etching, laminating, lithography, laser printing or others to generate pores, holes or other structures in the solid surface. This solid surface is then prepared for accepting conjugating capture molecules.
  • pore diameter and depth may be similar or may vary depending on the application. In some layers, pore sizes may vary, the variance of which may depend on the application.
  • Porosity that may be imparted on a Molecular Net may include but are not limited to picopores, nanopores, micropores, filtration pores, sieving pores, pockets or other. Porosity may be imparted into a Molecular Net by the selection of and method of incorporation of specific capture molecules, linker molecules and spacer molecules into each layer of a Molecular Net. Porosity may also be imparted into a Molecular Net by the selection of and method of incorporation of specific capture molecules, linker molecules and spacer molecules used in the fabrication of sequential layers.
  • Molecular Net porosity may range from about 6 Angstroms in diameter to more than about 1 um in diameter based on the identity of capture molecules, linker and spacers used in a layer.
  • the porosity of a Molecular Net may comprise a range of pore diameters. Exemplary diameter ranges may be from about 5 nm to about 50 nm, from about 10 nm to about 100 nm, from about 50 nm to about 200 nm, from about 250 nm to about 500 nm, from about 500 nm to about 1 um, and from about 800 nm to about 1.5 um.
  • capture molecules may be used to generate pores in a Molecular
  • capture molecules may be pre-linked to one another prior to being incorporated into a Molecular Net layer.
  • linkers may be selected based on Angstrom length of the spacer arms.
  • extenders may be used to connect a first linker to a second linker to generate a long multi-functional linker.
  • spacers may be used to generate pores in a Molecular Net. Spacers may also be pre-linked to one another prior to being incorporated into a Molecular Net layer.
  • inert physical plugs may be used to build a pore, whereby each physical plug may be placed on a previously built layer while a new layer is being constructed. After curing, the physical plugs may be removed, thus leaving a pore of a specific diameter.
  • a Molecular Net comprises a single type of capture molecule. In other examples, a Molecular Net comprises multiple types of capture molecules.
  • the use of more than one monoclonal antibody during the fabrication of a Molecular Net enables said Molecular Net to bind more than one epitope of an analyte.
  • Use of more than one type of epitope-specific capture molecule enables improved analyte capture by a Molecular Net and relates to its performance.
  • the use of more than one nucleic acid sequence may be used during the fabrication process to generate a Molecular Net capable of binding to more than one epitope of an analyte.
  • Examples of benefit depend on the use of the Molecular Net and may comprise improved performance in terms of minimum levels of detection, sensitivity, positive predictive value, negative predictive value, ability to work with degraded samples, ability to work with a diverse population, and others when used in a test; may comprise improved performance in binding capacity, purity, binding kinetics, target analyte depletion, and others when used as a purification tool; or other.
  • the use of capture molecules directed against mutually- confirmatory analytes may be used in a Molecular Net and relates to its performance.
  • Use of mutually-confirmatory capture molecules in a Molecular Net may be used in a confirmatory manner, whereby the capture of more than one analyte may provide a more statistically significant positive result; may provide a more robust test result; may provide additional information regarding a sample; and other.
  • Use of mutually-confirmatory capture molecules in a Molecular Net may also be used to qualify a sample or may be used as a control in a test or may be used to measure more than one related molecular variable linked to a disease state or may be used to measure more than one related molecular variable linked to the treatment of a disease.
  • Examples of mutually-confirmatory analytes a Molecular Net may be fabricated to simultaneously capture from a sample may include: genetic sequence and corresponding protein product (for examples, cancer-related SNPs in BRCAl and BRCAl protein); the mRNA and corresponding protein product (for example, human lactase mRNA and Lactase protein); the genetic sequence and the corresponding mRNA product (for example, disease-related SNPs in LMNA and pre-spliced or spliced Lamin A/C mRNA); miRNA and related mRNA or protein products (miR 9 and REST or CoREST mRNA, or miR 9 and REST protein); small molecule drugs and drug targets (tofacitinib and Janus kinase 3); epitope-specific biologies and the respective targets (for example, anti-TNF antibodies and circulating TNF cytokine); epitope- specific antibodies, epitope-specific T cells and/or epitope-specific B cells
  • Examples of benefit depend on use and may relate to improved performance in test sensitivity, positive predictive value, negative predictive value, specificity, diagnosis of a disease, ability to work with samples experiencing genetic drift, ability to measure response to a therapeutic, ability to measure effectiveness of a therapeutic, or other.
  • a Molecular Net may be fabricated in a manner to capture and position bound analytes in a manner that enhances the intensity of a detectable signal or may enhance detection of bound analytes, such as when used in a test with optical detection.
  • Placement of captured analytes in a layered manner by pre-positioned layered capture molecules may enable the rapid detection of analyte by signal intensification.
  • Examples of signal intensification by a Molecular Net may relate to fluorescence, fluorescence resonance energy transfer, absorbance, luminescence, light scatter, surface plasmon resonance, optical heterodyne detection, or other.
  • Said Molecular Net may be designed and fabricated to replace the need for costly and time-intensive methods for ultra-sensitive detection such as PCR, branched DNA, or multi- step detection methods required for signal amplification. Said Molecular Net may also be designed and fabricated to replace the need for costly and complicated analytical devices. [0030] Generally, the number of capture molecules incorporated into a 3-dimensional
  • Molecular Net matrix is less than or equivalent to the number of capture molecules conjugated in a 2-dimensional manner to a surface using conventional approaches.
  • Two-dimensional capture molecule-surface conjugates may rely on the use of a single linker type or may rely on the sequential use of 2 linkers to conjugate capture molecules to a solid surface.
  • multiple linker types are used simultaneously to link capture molecule to capture molecule of a new layer and the linked capture molecules of a new layer to a spacer or capture molecule of a previous layer.
  • Molecular Nets may be fabricated in solution prior to placement on a solid surface. Pre-fabricated Molecular Nets may be absorbed or covalently linked to a solid surface.
  • Molecular Nets may also be fabricated directly onto a solid surface, layer by layer. Said Molecular Nets may be placed on a solid surface using non- covalent (electrostatic, van der Waals, or other) or covalent methods.
  • polystyrene, polyurethane, polyethylene or treated surfaces such as poly-L-lysine coated surfaces, modified surfaces comprising -COOH, NHS, amine or other may be purchased from commercial sources (examples of vendors may include Thermo, Millipore, Luminex and other) and used as solid phase surfaces for Molecular Net placement.
  • solid phase surfaces may be pre-treated by chemicals such as acid to activate the surface moieties and thus to generate attachment points between the solid phase surface and reactive moieties of a Molecular Net.
  • a solid phase may be pre-treated with linker to covalently link a solid phase surface to a Molecular Net.
  • Design and fabrication of a Molecular Net for use on a solid phase surface may result in a covalently-linked multilayered three-dimensional matrix of capture molecules secured by covalent connectors within each layer.
  • Design and fabrication may occur in a sequential manner where a first layer is fabricated and subsequent layers are fabricated in a sequential manner whereby each layer may be interconnected in a covalent manner to enhance structural integrity, topology, porosity and/or stability.
  • Selection of individual capture molecules, linkers and spacers may be made to contribute to one or more property of a Molecular Net. Properties may comprise analyte specificity, thermal stability, layer thickness, pore diameter, absorbance spectra, emission spectra, solid phase compatibility or other.
  • topological features that may be imparted on a Molecular Net may include but are not limited to dimples, pocks, stipples, pores, mounds, branches, filaments, fibers, fissures, raised segments or other and may be arranged in a Molecular Net in a random, pseudorandom or irregular manner.
  • Topological features of a Molecular Net may be generated through the use of capture molecules and linkers; capture molecules, linkers and spacers; or linkers and spacers.
  • capture molecules may be used to generate topological features of a Molecular Net.
  • capture molecules may be pre-linked to one another prior to being
  • linkers may be selected based on Angstrom length of the spacer arms.
  • spacers may be used to connect a first linker to a second linker to generate a long multi-functional linker.
  • spacers may be used to generate topological in a Molecular Net. Spacers may also be pre-linked to one another or to capture molecules prior to being incorporated into a Molecular Net layer.
  • Molecular nets may be designed and fabricated to impart characteristics such as affinity, size exclusion, filtration, fluorescence, and other into each layer of a Molecular Net.
  • Specific capture molecules, linker molecules and spacer molecules may be selected based on size, length, diameter, thickness, optical properties, chemical properties or other for imparting characteristics into a Molecular Net during the fabrication process.
  • Molecular Nets may be fabricated in a manner whereby one or more capture molecules may serve a structural role, may serve both a structural role and a role in analyte capture within the covalently-linked multilayered three-dimensional matrix. Some examples of capture molecules that may be used for structural and/or analyte capture roles in a Molecular Net.
  • the distance between capture molecules in each layer of a Molecular Net may be determined, in part, by the diameter, width and/or length of capture molecules, linkers and spacers used in the fabrication process for each layer, whereby the molar relationship between each linker-capture-spacer molecule may be similar or may be different and the selection of said molecules may be dependent on size and/or shape of the analyte to be captured, the method used to measure captured analyte and/or desired use.
  • Molecular Nets may be designed and fabricated in a manner whereby each capture molecule, linker and spacer component may have equivalent or non-equivalent molar ratios in a layer of said Molecular Net. Variance of molar ratios between said components may be used from time to time to generate porosities or other topological features within each layer. Said porosities and topological features may have a range of diameters and may have a range of associated depths. Variance of molar ratios between Molecular Net components may occur in a single layer of Molecular Net or may occur in more than one layer of a Molecular Net and is dependent on the intended use of a Molecular Net.
  • analyte dimensions that may be considered during the design and fabrication process are provided in Table 2.
  • Design and fabrication of Molecular Net surface chemistry, pore diameter, topology, layering or other may be based on analyte shape; analyte structure, analyte isoforms, analyte charge, analyte complex formation with other molecules, and other forms.
  • Molecular Nets may be designed and fabricated to bind and capture said analyte or may be designed and fabricated to exclude said analyte. Examples of analytes and analyte sizes can be found in Table 2. TABLE 2. Examples of analytes and their dimensions.
  • Molecular Nets comprising structural components and capture components may be arranged in the covalently linked 3-dimensional (3D) multilayered matrix and may relate to the capture of one or more analyte relating to one or more of the following characteristics: surface chemistry; analyte shape; analyte structure; analyte isoforms; analyte charge; post- translational modification; chemical modification; activity; or other.
  • 3D 3-dimensional
  • Molecular Nets may comprise structural components that also act in a manner relating to the capture of analytes and may be arranged in the interconnected 3D multilayered matrix of a Molecular Net by covalent linkers.
  • a Molecular Net may also comprise spacers to interconnect said structure/capture molecules in a manner to maximize structural reinforcement, stability and/or specific analyte capture capability.
  • Molecular Net examples comprising capture components/structural components, linkers and spacers are presented in Table 3.
  • Klebsiella spp. Antibodies against Covalently linked Diagnostics Food surface antigens (e.g., antibodies - IgG, safety, infectious LPS, other); PNA IgM, covalent disease, water safety; probes against linkers, spacers Molecular tools: chromosomal and/or polymicrobial plasmid DNA sampling, microbiome sampling, molecular biology
  • Pseudomonas spp. Antibodies against Covalently linked Diagnostics Food surface antigens (e.g., antibodies - IgG, safety, infectious LPS, V antigen, other), IgM, covalent disease, water safety; excreted materials linkers, spacers Molecular tools: (e.g., heat shock polymicrobial proteins, alginate, sampling, microbiome other); PNA probes sampling, molecular against chromosomal biology
  • Staphylococcus Antibodies against Covalently linked Diagnostics Food aureus surface antigens (e.g., antibodies - IgG, safety, infectious protein A, IgM, covalent disease, water safety; peptidoglycan, other), linkers, spacers Molecular tools: excreted materials polymicrobial (e.g., heat shock sampling, microbiome proteins, exotoxins, sampling, molecular other); PNA probes biology
  • Staphylococcus Antibodies against Covalently linked Diagnostics Food aureus (cluster) surface antigens (e.g., antibodies - IgG, safety, infectious protein A, IgM, covalent disease, water safety; peptidoglycan, other), linkers, longer Molecular tools: excreted materials spacers polymicrobial (e.g., heat shock sampling, microbiome proteins, exotoxins, sampling, molecular other); PNA probes biology
  • IgM Antibodies antibodies, response profiling, against 5 IgM; antigens, covalent vaccination, antibody antigens linkers, spacers titering; Molecular tools: immunologic studies, pre-clinical studies
  • CD79a others; IgM, covalent disease monitoring; antigens; TCR:antigen; linkers, spacers, Molecular tools: MHC Lantigen; MCH MHC: antigen immunologic studies, II: antigen; cytokines complexes, pre-clinical studies (e.g., ILIO, IL6, TGFb, cytokines
  • CD79a others; IgM, covalent disease monitoring; antigens; TCR:antigen; linkers, spacers, Molecular tools: MHC Lantigen; MCH MHC: antigen immunologic studies, II: antigen; cytokines complexes, pre-clinical studies (e.g., ILIO, IL6, TGFb, cytokines
  • MHC Lantigen MHC Lantigen
  • MCH MHC antigen immunologic studies, II: antigen
  • cytokines complexes pre-clinical studies (e.g., ILIO, IL6, TGFb, cytokines
  • B cell - late S B cell - late S, G2 Antibodies against Covalently linked Diagnostics: Immune and M phases of PAX5, CD 19, CD20, antibodies - IgG, response profiling, cell cycle CD79a, others; IgM, covalent disease monitoring;
  • TCR antigen receptor
  • linkers linkers, spacers, Molecular tools:
  • MHC Lantigen MHC Lantigen
  • MCH MHC antigen immunologic studies, II: antigen
  • cytokines complexes pre-clinical studies (e.g., IL10, IL6, TGFb, cytokines
  • Macrophage Complement Covalently linked Diagnostics: Immune antibodies against antibodies - IgG, response profiling, mannose receptor Ab, IgM, covalent infectious disease anti-Ly6C, or other; linkers, spacers, monitoring;
  • M2a, M2b, M2c complexes, monitoring; chronic markers; TLR agonists; cytokines, TLR inflammatory disease cytokines; DAMPs; agonists, DAMPs, monitoring; Molecular PAMPs; alarmins PAMPs, alarmins tools: immunologic studies, pre-clinical studies
  • neutrophil markers neutrophil markers
  • MHC antigen vaccination
  • TLR agonists complexes, monitoring; chronic cytokines; DAMPs; cytokines, TLR inflammatory disease PAMPs; alarmins agonists, DAMPs, monitoring; Molecular
  • PAMPs, alarmins tools immunologic studies, pre-clinical studies
  • Cytokines Antibodies against one Covalently linked Diagnostics Immune or more epitope of antibodies - IgG, response profiling, cytokine; cytokine IgM, covalent infectious disease binding domain of linkers, spacers, monitoring;
  • the solid phase may be particles ranging from about 2 nm in diameter to about 200 mm in diameter and Molecular Nets may be attached to the surface of said particle.
  • Particles may comprise polystyrene, polyethylene, silica, composite, nylon, PVDF, nitrocellulose, cellulosic, carbon, or other may be magnetic, paramagnetic, fluorescent, barcoded or other.
  • Molecular Nets may be absorbed or covalently linked to the surface of a particle in manner to generate pseudorandom or ordered porosities in a single layer of said Molecular Net or throughout.
  • a particle may be initially coated with a layer of Molecular Net, which may be connected to a second layer, which may be connected to a third layer.
  • Molecular Net layers may comprise the same capture molecules at the same or at different concentrations in each layer.
  • Molecular Net particles may also comprise different capture molecules in each layer and may be fabricated in a manner to incorporate the same or different concentrations of capture molecules compared to previous layers.
  • Molecular Nets may be attached to a particle surface in a manner to generate an asymmetric particle having a pre-determined polarity.
  • a particle may be designed and fabricated with an initial layer comprising structural molecules with a large diameter, width and/or length and may be linked to a particle in an asymmetric manner to generate a polarity.
  • a second layer may be linked to a first layer and a third layer may be connected to a second layer and so on.
  • the number of layers in a Molecular Net particle may vary depending on use.
  • Molecular Nets may be attached to a segment of a particle to generate an asymmetric particle having a pre-determined polarity.
  • a particle is constructed whereby the initial layer is coated onto a segment of a particle and whereby a second layer is connected to the initial layer onto the same segment of said particle, and whereby a third layer is connected to the second layer onto the same segment of said particle, and whereby a fourth layer is connected to the third layer onto the same segment of said particle.
  • Molecular Nets may be passively absorbed to a
  • particle surfaces may be functionalized and may require activation prior to attachment.
  • particle surfaces may be activated prior to functionalization, at which time a Molecular Net may be attached.
  • Molecular Nets may be constructed directly on the particle surface. Attachment of Molecular Nets to a particle may change the physical and/or chemical features of said particle.
  • Molecular Nets may comprise pseudorandom topological features placed on the surface of a particle.
  • Molecular Net particles may comprise topological features, the topological features comprising capture molecules and linkers and may also comprise spacers.
  • topological features may include appendages, spikes, plateaus, planes, mounds, fissures, pellicles, stipples, channels, pores and other and may be comprised of capture components directly linked within and/or linked to one or more layer of a Molecular Net.
  • topological features may comprise be pockets, pillars, bumps, branches, projections, ridges, clefts, trellis-like structures, flakes, pellets, spheres, or others. Topological features may be pre-formed in solution and linked to the Molecular Net or may be formed at the time each layer is constructed.
  • Molecular Nets on particles may comprise heterogeneous capture molecules within one or more layer of a Molecular Net.
  • Benefits of a heterogeneous design may relate to the capture of a plurality of analytes having a plurality of surface chemistries on a single particle.
  • Heterogeneous capture molecules incorporated into a Molecular Net during fabrication may be randomly distributed throughout each layer; may be stratified throughout each layer; or other, depending on use.
  • Molecular Nets may be attached to particles to increase surface area of said particle. Molecular Nets may also be used to increase particle diameter. Topological features of a Molecular Net on a particle may relate to an increased particle size in addition to analyte capture capacity.
  • a first layer of Molecular Net may be attached to a particle surface to modify the physical and/or chemical properties of particle.
  • “bead effects” or “surface effects” can hamper results and are still not well understood.
  • Traditional conjugation techniques that result in 2D conjugates and 2D conjugated surfaces often suffer from surface effects.
  • Molecular Nets may be used to minimize or neutralize bead effects to minimize non-specific binding to a bead surface, bead autofluorescence, bead interference with in an assay or other.
  • Molecular Net particles may impart increased analyte binding capacity and may also impart blockade of non-specific binding of undesired analytes to increase the signal-to-noise ratio in an assay, yield and purity of purified analyte or other.
  • the invention features molecular Net on particle containing more than one layer, wherein each layer contains capture molecules directed against analyte, wherein each layer contains distinct capture molecules directed against distinct analyte, wherein different layers can be directed against different analytes to enable the capture of analyte or a plurality of analytes.
  • a Molecular Net placed on a solid phase surface may be used to increase the purity of one or more analyte recovered from a sample.
  • Molecular Net- coated surfaces may reduce non-specific binding of undesired analyte compared to commercial 2D functionalized surfaces.
  • Molecular Nets placed on particles may significantly increase analyte capture capacity of a particle. Additional layering of a Molecular Net may further increase the number of bound analyte per particle and may be used to enhance recovery or yield of analyte from a sample and may be used to deplete one or more analyte from a sample.
  • Molecular and cellular testing strategies employ the use of single-plex or multiplex immunoassays, PCR assays, next-generation sequencing techniques or other to identify the presence of or to measure the amount of one or more analyte in a sample.
  • reactions may be separated spatially or may be combined into a single testing reaction and may employ solid phases comprising unique identifiers to provide information.
  • unique identifiers may comprise the use of different barcodes, different fluorescence emissions, different chemistries, different ordered nucleotide tags, or other.
  • Solid phases may be used in single-plex and multi-plex assays may rely on the specific binding of target analyte to produce a measurable signal or a measurable change in signal and may be used in a direct assay or may be used in an indirect assay.
  • Measurable signals may be generated from a positive test and may comprise electric, thermal, magnetic, optical, vibrational, isotopic, or other measurable characteristic.
  • Molecular Nets may be used in place of current commercial approaches and may generate specific and sensitive analyte capture, detection and measurement from a sample.
  • Advantages of a Molecular Net are presented in Table 4 and may include: the rapid capture of one, several or a plurality of molecular and cellular analytes in a raw sample; ability to generate sensitive and specific signals when used in a test involving indirect and direct detection methods; ability to generate a signal having enhanced fluorescent intensity; ability to concentrate bound analyte; ability to spatially separate bound analyte in a manner that reduces stearic hindrance between analytes and/or between detection molecules; enhanced stability; reduced background and others.
  • Stability of test (enzyme-free) longer Increases shelf life, & reduces costs associated shelf life with storage issues
  • Molecular Nets may be used in applications where analyte binding efficiency, analyte binding kinetics, analyte binding capacity, analyte detection, analyte measurement, analyte enrichment, analyte purification and analyte delivery may be important.
  • Molecular Nets may be used in fluid phase or may be attached to a solid phase.
  • Molecular Nets may be attached through absorption or covalent processes on a receptive surface.
  • solid phases include but are not limited to nanotubes, metals, particles, microtiter plates, slides, cassettes, probes, lateral flow tests, stents, catheters, valves, blood tubes, needles, solid phase devices or other.
  • chemistries of various solid phases that may be compatible for Molecular Net attachment include but are not limited to plastics, other polymer, thin film, colloidal metals, silica, carbon nanotube, protein,
  • Molecular Nets may be attached to a solid phase device surface to capture, purify or deplete one or more analyte from a sample.
  • An example of using a Molecular Net for analyte capture and/or purification from a sample is presented in Figure 1.
  • Molecular Nets that may be used to capture and purify analyte from a sample are: Protein A, Protein G or Protein L Net-coated microspheres for immunoglobulin capture; Streptavidin Net- coated microspheres for biotin capture; TNF Net-coated microspheres for anti-TNF biologic capture; IL6 Net-coated microspheres for anti-IL6 biologic capture; IgM Net-coated microspheres for RNA virus capture; Ig Fc Net-coated microspheres for complement capture; antigen Net-coated microspheres for antigen-specific immunoglobulin; antigen-specific immune cell capture; and others. Molecular Nets may be used in chromatography methods for the capture and purification of one or more analyte from a sample.
  • Molecular Nets may be used to capture analyte from a sample for downstream analyte measurement by an independent method, referred to herein as sample prep.
  • independent methods may include mass spectrometry, immunoassay, PCR, next-generation sequencing, qRT-PCR, digital PCR, microscopy, fluorescence, flow cytometry, bead cytometry, or other.
  • the invention improves signal-to-noise ratios when used in an assay.
  • Molecular Nets may be attached to a solid phase device surface to measure the presence, absence, modification or concentration of one or more analyte. Examples of using Molecular Nets for analyte detection and/or measurement are presented in Figures 3 and 4. In some other examples, Molecular Nets may be used to simultaneous detect and measure 2 or more specific analytes in a direct or indirect manner. Indirect capture by a Molecular Net may relate to the capture of a primary analyte by a specific capture molecule of a Molecular Net that may enable the detection of one or more related secondary associated with the captured primary analyte.
  • Molecular Nets may be used as discovery tools capture primary analytes from a sample and enables the identification, detection or measurement of secondary analytes that are captured- by-association. Molecular Nets may be used in this manner for drug discovery, pathway mapping, and in proteomics, transcriptomics, glycomics, lipidomics, metabolomics, functional genomics, foodomics, nutrition, pharmacology, toxicology and others.
  • Molecular Nets may be used to detect drug resistance in a cell.
  • Cells may be tumor cells, immune cells, microbial cells or other cells.
  • Molecular Nets for these applications may comprise capture molecules directed against one or more unique features of a cell type.
  • Molecular Nets may additionally be fabricated in a manner to impart surface topology relating to the capture of in tact cells.
  • Molecular Nets may be fabricated and used in: immune cell reactivity measurement; immune response monitoring; immune response classification;
  • immunoglobulin titering biotinylated molecule capture; multiplex immunoassays; singleplex immunoassays; next-generation sequencing reactions; PCR; microbiome capture; microbiome discovery; mRNA and encoded protein measurement; SNP (single nucleotide polymorphisms) mapping; SNP detection; disease marker sample preparation; miRNA capture and/or
  • Molecular Nets may have measurable characteristics imparted during the fabrication process and may be used in a direct or indirect manner as a sensor.
  • a measurable change in one or more characteristic of a Molecular Net sensor may be detected using commercial approaches employing the use of optical sensing, electrochemical sensing, electromagnetic sensing, electrical impedence, or other.
  • a Molecular Net sensor may be used to capture and bind an analyte. Analyte binding may result in a measurable change in a characteristic of a Molecular Net sensor.
  • a binding event or modifying event pertaining to the Molecular Net sensor may be monitored over a period of time, and the changes in Molecular Net sensor characteristics may be detected, relayed and collected by a device.
  • Other examples of using a Molecular Net as a sensor may include an analyte binding event, enzymatic reaction, analyte modification event, cell differentiation, cell-cell interaction, or other.
  • measurable characteristics include but are not limited to: physical shape, height, density, fluorescence intensity, wavelength shift (FRET or FRAP), vibrational frequency, absorbance, flexibility, refractiveness, conductance, impedence, resistance, melting temperature, denaturation temperature, freezing temperature, and other.
  • Measuring devices that may be compatible for use with a Molecular Net sensor may comprise: photonic multichannel analyzers, spectrometers, magnetic resonance imagers, magnetic field detectors, optical fibers, glass pipettes, circuits, fluorometers, spectroscopic analyzers, flow cytometers, CCD cameras, microscopes, acoustic chambers, microphones, luminometers, and other.
  • the measuring devices may be used to measure changes in: thickness, topology, charge, insulation, capacitance, voltage, color, acoustics, vibration, magnetism, enzymatic activity or other characteristics of a Molecular Net used as a sensor.
  • Molecular Nets may be used in flexible circuits, whereby the capture molecules and/or structural molecules may be connected to conductive molecules.
  • Molecular Net circuits may be used in single-sided flexible circuits, double access (back bared flex circuits), sculptured flex circuits, double-sided flex circuits, multilayered flex circuits, ridge flex circuits, ridge-flex boards, polymer thick film flex circuits or other.
  • Most flexible circuits are passive wiring structures that are used to interconnect electronic components such as integrated circuits, resistor, capacitors and the like, however some are used only for making interconnections between other electronic assemblies either directly or by means of connectors.
  • Molecular Nets for use in circuits or for use as a component of a circuit may be comprised of synthetic components or may be comprised of biochemical capture molecules and/or cells and may be fabricated in a manner to be used in a flexible circuit.
  • Molecular Net circuits may also be used as a sensor.
  • Molecular Net circuits may have specific electrochemical properties and may be used to monitor various parameters such as pH, current, voltage, impedence, or other in an electrochemical/electrolyte cell. Binding events and modifying events that may occur to Molecular Net may be measurable and may be reflected by a change in the conductance, current, or voltage. More specifically, the introduction of a sample containing an analyte that has specific binding affinity for, or is reactive towards, a component in a Molecular Net circuit may be monitored by a change in the electrochemical properties of the Molecular Net and/or the surrounding environment.
  • binding events on a Molecular Net used in a circuit may include: antibody-antigen interaction, nucleic acid-nucleic acid interaction, enzyme-substrate interaction, drug-target interaction, enzyme-co-factor interaction, ligand-cell interaction, or any other specific surface-chemistry-driven non-covalent interaction.
  • Analyte capture by a Molecular Net may be determined by a change in pH, current or voltage an electrochemical/electrolyte cell. Measurement of a change in Molecular Net characteristics may also result from one or more, or an accumulation of modifying events to one or more component in a Molecular Net or to a capture analyte.
  • modifying events may include: enzyme cleavage; post- translational modification (such as phosphorylation, sulfonation, glycosylation, methylation, or other); removal of a post-translational modification (such as de-phosphorylation); or other similar modification.
  • Modifying events may be determined by a change in pH, current or voltage in the electrochemical/electrolyte cell resulting from a change in Molecular Net characteristics or in the surrounding buffer system.
  • Methods to determine changes in electrochemical properties of a Molecular Net used in a circuit may include the use of scanning ion current microscopy, nanofluidic diodes, nanopores or nanochannels that display voltage-gated ion current, ion nanogating, nanopore- based sensing platforms and other methods for measuring the flow, or changes in flow of electrical charge through a medium. More specifically, the inherent sensitivity of many solid- state nanopore sensors is the selective permeability of electrolytes, or ion current, when a bias is applied across the nanopore. Molecular Nets may be coated onto the surface of a nanopore and the change in current, voltage, and impedance can be monitored.
  • Molecular Net can also be coated onto the surface of a carbon nanotube and whereby the molecular Net can be constructed in a manner to generate size exclusion and affinity requirements for analyte sensing.
  • Bio-Layer Interferometry is a label-free technology for measuring biomolecular interactions. It is an optical analytical technique that analyzes the interference pattern of white light reflected from two surfaces: a layer of immobilized molecular Net on the biosensor tip, and an internal reference layer. Any change in the number of molecules bound to the biosensor tip causes a shift in the interference pattern that can be measured in real-time.
  • the binding between a ligand immobilized on the molecular Net-coated biosensor tip and an analyte in solution produces an increase in optical thickness at the biosensor tip, which results in a wavelength shift, ⁇ , which is a direct measure of the change in thickness of the biological layer.
  • Interactions may be measured in real time, providing the ability to monitor binding specificity, rates of association and dissociation, or concentration, with precision and accuracy. Only molecules binding to or dissociating from the Molecular Net biosensor may shift the interference pattern and generate a response profile. Unbound molecules, may change the refractive index of the surrounding medium, or may change flow rate but will not affect the interference pattern. This is a unique characteristic of BLI and extends its capability to perform in crude samples used in applications for analyte - capture molecule binding, quantitation, affinity, and kinetics.
  • Molecular Net particles may also be used to deliver an active agent.
  • Active agents may be pre-loaded onto capture molecules located in one or more layer of a Molecular Net.
  • Active agents may comprise: drugs, therapeutics, toxins, viruses, allergens, vaccine components, antigens, immune modulators, surfactants, microbes, oligonucleotides, nutrients, or other.
  • Molecular Net particles may be used in drug or therapeutic delivery, vaccine delivery, in biofermentation or other.
  • Molecular Nets may comprise one or more targeting agent on a surface-exposed layer to facilitate specificity in targeting said Molecular Net particle to a specific cell type, tissue type, organ type or other.
  • Targeting agents may be capture molecules of a Molecular Net.
  • Targeting agents may comprise: antibodies, receptors, ligands, anti-ligands, or other.
  • Targeting agents in a Molecular Net may be covalently linked to capture molecules, linkers, and spacers in a surface-exposed layer. Targeting agents may also contribute to the topological features of a Molecular Net.
  • microparticles for analyte purification are provided.
  • Molecular Nets comprised of monomeric protein G and linked protein G and crosslinkers BS 3 , EMCS, EGS, BMPH and others were used in fabrication. Molecular Net fabrication occurred in real-time on 0.8-10 um magnetic polystyrene microparticle and 45 um nitrocellulose microparticle surfaces.
  • the capture molecule, protein G was used as the only source of structural support.
  • pre-linked protein G and monomeric protein G were mixed to serve as additional structural support for fabrication of some layers of the Molecular Net.
  • a first layer of Molecular Net comprised protein G and Ig Fc region to serve as structural support for fabrication of additional layers of the Molecular Net.
  • a protein G Molecular Net comprised 2 layers and in other examples, a protein G Molecular Net comprised 3 layers.
  • Figure 1 is an example of data obtained using protein G Molecular Net microparticles in comparison to commercial protein G microparticles. Briefly, IgG-Alexa 647 was spiked into human serum (1 ug/tube). Uncoated microparticles, commercial protein G microparticles and protein G Molecular Net microparticles were incubated with spiked sample for 15-60 min at RT (at 100,000 particles (uncoated control), 100,000 particles (commercial) and 25,000 particles (Molecular Net).
  • Particles were then quenched, blocked, and washed prior to use. Particles were incubated for 15 min in pre-cleared serum + neuroserpin at a concentration range of (0-1 ng/mL). Bound particles were washed and incubated with biotin-anti-Neuroserpin (10 ng/mL) for 15 min. Neuroserpin detection was visualized by avidin-PE (30 ng/mL) for 15 min. Washed particles were then analyzed on a Luminex 100, collecting 100 particles per sample. Presented in Figure 2 is the median fluorescence intensity (FI) at each dilution above background fluorescence intensity.
  • FI median fluorescence intensity
  • FIG. 3 Effectiveness of Molecular Net microparticles in measuring analyte.
  • FIG. 4 Effectiveness of a Molecular Net with topology in measuring analyte.
  • Molecular Net comprised of identical anti-human neuroserpin antibody (as
  • FIG. 5 Comparison of traditionally conjugated microparticles and Molecular Net microparticles in an ELISA.
  • Molecular Nets comprised of monoclonal antibody directed against human-TauF and crosslinkers Sulfo-NHS, EMCS, EGS, BMPH and others were used in fabrication.
  • the capture molecule, anti-Tau mAb was used as the only source of structural support.
  • the spacer, albumin was mixed with the anti-Tau mAb in a first layer at a 1.5: 1.0 Molar ratio (albumin:anti-Tau Ab) to serve as additional structural support for fabrication of the first layer.
  • a second capture molecule, human tubulin was used and provided both structural support and capture roles within a Molecular Net.
  • Figure 5 is an example of data obtained using an anti-Tau Molecular Net (LV.P6 Cap-TECH) in comparison to a commercial Tau microparticle (Partner LV) ELISA (identical assay conditions, identical antibody pair, etc.).
  • Figure 5 is an example of using a Molecular Net to reduce background noise in an assay and increase a visible signal in an ELISA.
  • FIG. 6 Comparison of traditionally conjugated microparticles and Molecular Net microparticles using Luminex.
  • Molecular Net comprised of monoclonal antibody directed against human-thyroid stimulating hormone and crosslinkers EDC, BS(PEG)g, EMCS, EGS, BMPH and others were used in fabrication. Molecular Net fabrication occurred in real-time on Luminex magnetic microparticle surfaces.
  • the capture molecule, anti-TSH mAb was used as the only source of structural support.
  • the spacers, PEG, heat-denatured lysozyme and others were mixed with the anti-TSH mAb in a first layer at a 1.0:2.0 Molar ratio
  • an anti-TSH Molecular Net comprised 4 layers and in other examples, an anti- TSH Molecular Net comprised 6 layers with the last layer comprising topological features to enhance analyte binding and performance in a Luminex.
  • Figure 6A is an example of using a Molecular Net to increase the overall MFI in a Luminex assay.
  • Figure 6B is exemplary data obtained in a Luminex assay to increase the minimum levels of detection in a Luminex assay.
  • FIG. 7 Exemplary Molecular Nets on particles.
  • Figure 7 depicts some examples in which Molecular Nets may be placed onto a particle surface.
  • a Molecular Net is placed on a particle surface (Fig. 7A, 1001) in a circumferential manner whereby a Molecular Net having X, Y and Z spatial orientation may be fairly symmetrical and where each layer (examples of 3 layers, 1002, 1003 and 1004) adds to the Z plane of the particle.
  • Molecular Nets may be placed onto a particle surface (Fig. 7A, 1001) in a circumferential manner whereby a Molecular Net having X, Y and Z spatial orientation may be fairly symmetrical and where each layer (examples of 3 layers, 1002, 1003 and 1004) adds to the Z plane of the particle.
  • Molecular Nets may be placed onto a particle surface (Fig.
  • Molecular Nets may be placed onto a portion of particle surface (Fig.
  • FIG. 8 Exemplary Molecular Net topological features on particles.
  • Figure 8 depicts some examples in which Molecular Nets may be placed onto a particle surface.
  • a Molecular Net is placed on a particle surface (Fig. 8A, 2001) in a circumferential manner whereby a Molecular Net having X, Y and Z spatial orientation may be fairly symmetrical in one layer (2002) and where each layer (examples of 3 layers, 2002, 2003 and 2004) adds to the Z plane of the particle in differing and asymmetric ways (for example, topology is generated).
  • Molecular Nets may be placed onto a particle surface (Fig.
  • a Molecular Net having X, Y and Z spatial orientation may be placed onto the surface of a particle in a manner to generate structural features (2008) throughout layers 1 (2006), 2 (2007) and 3 (2008) of the particle and where each layer adds to the Z plane of the particle in a layer-dependent manner (for example, some layers may have less height and other layers may have more height).
  • each layer may also serve an analyte capture role in a Molecular Net.
  • Molecular Nets may be placed onto a portion of particle surface (Fig. 8C, 1009) in an asymmetrical manner whereby a Molecular Net having X, Y and Z spatial orientation may be placed onto the surface of a particle in a manner to generate a polarity of the particle and where each layer (examples of 4 layers, 2010, 2011, 2012 and 2013) adds to the Z plane of the particle in a layer-dependent manner and whereby each layer may serve both structural and analyte capture roles.
  • some layers may have specificity for an analyte based on size (e.g., Fig. 8C, 2010) and outer layers (e.g., Fig. 8C, 2013) may have specificity for analyte of a larger size.
  • FIG. 9 Exemplary Molecular Nets for analyte delivery.
  • Figure 9 depicts some examples in which Molecular Nets may be placed onto a particle surface for use in analyte capture or targeted analyte delivery.
  • a Molecular Net is placed on a particle surface (Fig. 9A, 3001) in a circumferential manner whereby a Molecular Net having X, Y and Z spatial orientation may be fairly symmetrical in one layer (3002) and where each layer (examples of 3 layers, 3002, 3003 and 3004) adds to the Z plane of the particle in differing and asymmetric ways (for example, topology is generated).
  • analyte cargo (3003) may be pre-loaded onto capture molecules in one or more layer of a Molecular Net.
  • a pre-loaded analyte may comprise a drug, a therapeutic, siRNA, miRNA, dsRNA, virus, toxin, immunogen or other.
  • Pre-loaded cargo may be non-covalently associated with one or more type of capture molecule in a layer of a Molecular Net.
  • different capture molecules (3004) of a Molecular Net may be arranged in the outer layers of a Molecular Net and may serve topological and analyte capture roles.
  • the different capture molecules may have specificity for one or more different analyte, the analyte of which may comprise an antibody, an anti-ligand, a ligand, a receptor, an antigen or other and may serve one or more structural and/or affinity and/or targeting role.
  • analyte of which may comprise an antibody, an anti-ligand, a ligand, a receptor, an antigen or other and may serve one or more structural and/or affinity and/or targeting role.
  • Molecular Nets may be placed onto a particle surface (Fig. 9B,
  • analyte cargo (3006) may be pre-loaded in all layers of a Molecular Net.
  • capture molecules may be used to generate topological features (3008) that serve particle-targeting roles.
  • outer layers of a Molecular Net particle may target said particle to a specific cell, tissue, organ or other.
  • FIG. 10 Molecular Nets for Analyte Purification from a Sample
  • Molecular Nets may be used in sample purification processes (example is provided in Figure 10).
  • Molecular Net designed and fabricated to deplete one or more analyte may be used to treat a sample for analyte depletion.
  • Exemplary methods may include incubation of Molecular Net with sample for about 15 minutes to about 24 hours in a batch slurry or in a chromatography column. Sample supernatant or flowthrough may be collected depending on preferred method.
  • Molecular Nets may be collected and analyzed using various methods for the presence and amount of captured analyte.
  • Molecular Net-treated sample may be collected and analyzed using various methods to determine the residual presence of analyte in the sample or may be analyzed for other analytes in the sample.
  • FIG. 11 Molecular Nets for Analyte Detection & Measurement from a Sample
  • Molecular Nets may be used in an analyte measurement tool or a diagnostic tool
  • Molecular Net designed and fabricated to capture one or more analyte may be used to treat a sample for analyte detection and
  • Exemplary methods may include incubation of Molecular Net with sample for about 15 minutes to about 2 hours in a batch slurry, cassette, slide, microtiter plate or other. Sample supernatant or flowthrough may be collected depending on preferred method. Molecular Nets may be collected and analyzed using various methods for the presence and amount of captured analyte(s). Molecular Net-treated sample may also be collected and analyzed using various methods to measure other analytes. Methods for measuring changes in Molecular Net characteristics may include optical, electrophoretic, electrical, magnetic, chemical, thermal or other.

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Abstract

L'invention concerne une matrice tridimensionnelle multicouche liée par covalence comprenant des molécules de capture, des lieurs et des espaceurs (appelés filets moléculaires) pour une capture spécifique et sensible d'analyte d'un échantillon. L'invention concerne également un filet moléculaire comprenant une matrice tridimensionnelle multicouche liée par covalence comprenant plus d'un type de molécules de capture et plus d'un type de lieurs et pouvant comprendre un ou plusieurs espaceurs pour la capture spécifique et sensible de plus d'un type d'analytes d'un échantillon. Un filet moléculaire peut comprendre une nature pseudo-aléatoire. L'utilisation de diverses molécules de capture, de divers lieurs et espaceurs dans un filet moléculaire peut conférer des propriétés de liaison uniques à un filet moléculaire. La porosité, l'affinité de liaison, les capacités d'exclusion de taille, les capacités de filtration, les capacités de concentration et les capacités d'amplification de signal d'un filet moléculaire peuvent varier et dépendent de la nature des composants utilisés dans sa fabrication. Les utilisations d'un filet moléculaire peuvent comprendre la capture d'analyte, l'enrichissement d'analyte, la purification d'analyte, la détection d'analyte, la mesure d'analyte et la distribution d'analyte. Le filet moléculaire peut être utilisé dans une phase liquide ou sur des phases solides, telles que des nanomatériaux, des surfaces métalliques modifiées, des nanosphères, des microsphères, des plaques de microtitrage, des lames, des pipettes, des cassettes, des cartouches, des disques, des sondes, des dispositifs à écoulement latéral, des dispositifs de microfluide, des fibres optiques et d'autres.
PCT/US2014/029823 2013-03-14 2014-03-14 Filets moléculaires sur phases solides WO2014153262A1 (fr)

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