Devices for capturing analyte
Disclosed is a covalently-linked multilayered three-dimensional matrix comprising capture molecules, linkers and spacers (referred to as a Molecular Net) for specific and sensitive analyte capture from a sample. Also disclosed herein is a Molecular Net comprising covalently-linked multilayered three-dimensional matrix comprising more than one type of capture molecule and more than one type of linker and may comprise one or more spacer for specific and sensitive capture of more than one type of analyte from a sample. A Molecular Net may comprise a pseudorandom nature. Use of various capture molecules, linkers and spacers in a Molecular Net may confer unique binding properties to a Molecular Net. Porosity, binding affinity, size exclusion abilities, filtration abilities, concentration abilities and signal amplification abilities of a Molecular Net may be varied and depend on the nature of components used in its fabrication. Uses of a Molecular Net may include analyte capture, analyte enrichment, analyte purification, analyte detection, analyte measurement and analyte delivery. Molecular Nets may be used in liquid phase or on solid phases such as nanomaterials, modified metal surfaces, nanospheres, microspheres, microtiter plates, slides, pipettes, cassettes, cartridges, discs, probes, lateral flow devices, microfluidics devices, microfluidics devices, optical fibers and others.
1. A device for capturing an analyte, the device comprising:
a solid phase; and
a molecular net coupled to at least a portion of a surface of the solid phase, the molecular net comprising two or more layers, which two or more layers comprise at least a first layer and a second layer, and each of the two or more layers comprises capture molecules of at least one type capable of specifically binding to the analyte, which capture molecules are coupled to each other by linker molecules of a plurality of types,
wherein
the molecular net is a covalently-linked three dimensional matrix; and
at least the first layer and the second layer have porosities that are different from each other.
2. The device of claim 1 , wherein the solid phase is made of one or more of a plastic, polymer, thin film, colloidal metal, silica, carbon nanotube, protein, carbohydrate, lipid, nucleic acid, cell, or tissue.
3. The device of claim 1 , wherein the solid phase comprises one or more of a nanomaterial, modified metal surface, nanosphere, microsphere, microtiter plate, slide, pipette, cassette, cartridge, disc, probe, lateral flow device, microfluidics device, or optical fiber.
4. The device of claim 1 , wherein the molecular net is prefabricated and absorbed to the surface of the solid phase.
5. The device of claim 1 , wherein the molecular net is covalently linked to the surface of the solid phase.
6. The device of claim 1 , wherein the molecular net is constructed directly on the surface of the solid phase.
7. The device of claim 1 , wherein the capture molecules of the first layer, the capture molecules of the second layer, or both, comprise one or more of antibodies, nucleic acid probes, enzymes, recombinant proteins, or peptides.
8. The device of claim 1 , wherein the capture molecules of the first layer, the capture molecules of the second layer, or both, comprise a plurality of monoclonal antibodies for binding a plurality of epitopes of the analyte.
9. The device of claim 1 , wherein the capture molecules of the first layer, the capture molecules of the second layer, or both, are directed against mutually-confirmatory analytes.
10. The device of claim 1 , wherein the linker molecules of the first layer, the linker molecules of the second layer, or both, comprise one or more of homobifunctional, heterobifunctional, trifunctional, or multifunctional types.
11. The device of claim 1 , wherein the molecular net has at least one measurable characteristic that undergoes a change when the capture molecules of the first layer, the capture molecules of the second layer, or both, bind to the analyte.
12. The device of claim 11 , wherein the measurable characteristic comprises one or more of physical shape, height, density, fluorescence intensity, wavelength shift, vibrational frequency, absorbance, flexibility, refractiveness, conductance, impedance, resistance, melting temperature, denaturation temperature, or freezing temperature.
13. The device of claim 1 , wherein the capture molecules of the first layer, the capture molecules of the second layer, or both, are also coupled to each other by a plurality of types of spacer molecules.
14. The device of claim 13 , wherein the spacer molecules comprise one or more of PEGs, polymers, nucleic acids, albumins, Fc regions, or peptides.
15. The device of claim 13 , wherein the spacer molecules and an amount of spacer molecules are selected to give the molecular net one or more desired physical properties.
16. The device of claim 15 , wherein the desired physical properties comprise one or more of porosity, charge profile, or topological features.
17. The device of claim 1 , wherein the molecular net comprises 2, 3, 4, 5, or 6 layers.
18. The device of claim 1 , wherein each layer of the molecular net comprises at least one topological feature Which affects the porosity of the layer.
19. The device of claim 18 , wherein the at least one topological feature comprises one or more of fissures, channels, pores, or pockets.
20. The device of claim 1 , wherein the solid phase comprises an underlayer and the molecular net is coupled to at least a portion of the underlayer.
21. The device of claim 20 , wherein the underlayer does not comprise capture molecules, which underlayer capture molecules are capable of binding to the analyte.
22. The device of claim 1 , wherein the first layer is closer to the solid phase than the second layer, and the first layer has a first porosity that is less than a second porosity of the second layer.
23. The device of claim 1 , wherein the last constructed molecular net layer has a porosity that is greater than porosity of any other molecular net layer.
24. The device of claim 1 , wherein porosity increases from the innermost layer of the molecular net to the outermost layer of the molecular net.