WO2015168653A1 - Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material - Google Patents
Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material Download PDFInfo
- Publication number
- WO2015168653A1 WO2015168653A1 PCT/US2015/028948 US2015028948W WO2015168653A1 WO 2015168653 A1 WO2015168653 A1 WO 2015168653A1 US 2015028948 W US2015028948 W US 2015028948W WO 2015168653 A1 WO2015168653 A1 WO 2015168653A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- filter
- fluid
- membranes
- perforated
- membrane
- Prior art date
Links
- 239000012528 membrane Substances 0.000 title claims abstract description 190
- 239000000463 material Substances 0.000 title claims abstract description 98
- 238000000926 separation method Methods 0.000 title claims description 44
- 238000003556 assay Methods 0.000 title description 36
- 238000001914 filtration Methods 0.000 title description 25
- 239000012530 fluid Substances 0.000 claims abstract description 121
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 99
- 229910021389 graphene Inorganic materials 0.000 claims abstract description 85
- 239000011148 porous material Substances 0.000 claims abstract description 75
- 238000000034 method Methods 0.000 claims abstract description 51
- 239000000126 substance Substances 0.000 claims abstract description 25
- 230000007423 decrease Effects 0.000 claims abstract description 13
- 238000012986 modification Methods 0.000 claims description 9
- 230000004048 modification Effects 0.000 claims description 9
- 231100000765 toxin Toxicity 0.000 claims description 7
- 239000003053 toxin Substances 0.000 claims description 6
- 238000004891 communication Methods 0.000 claims description 2
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 230000002934 lysing effect Effects 0.000 claims description 2
- 230000009089 cytolysis Effects 0.000 claims 1
- 238000011002 quantification Methods 0.000 abstract description 2
- 238000012797 qualification Methods 0.000 abstract 1
- 238000004458 analytical method Methods 0.000 description 28
- 238000007306 functionalization reaction Methods 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 18
- 230000008901 benefit Effects 0.000 description 16
- 229910052799 carbon Inorganic materials 0.000 description 14
- 108090000623 proteins and genes Proteins 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 239000010410 layer Substances 0.000 description 11
- 230000007246 mechanism Effects 0.000 description 11
- 241000700605 Viruses Species 0.000 description 10
- 230000008859 change Effects 0.000 description 9
- 230000007547 defect Effects 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 8
- 230000006870 function Effects 0.000 description 8
- 108020004707 nucleic acids Proteins 0.000 description 8
- 102000039446 nucleic acids Human genes 0.000 description 8
- 150000007523 nucleic acids Chemical class 0.000 description 8
- 239000003575 carbonaceous material Substances 0.000 description 7
- 241000894007 species Species 0.000 description 7
- 239000013078 crystal Substances 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 108700012359 toxins Proteins 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 125000004429 atom Chemical group 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000013060 biological fluid Substances 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 230000004913 activation Effects 0.000 description 3
- 239000012620 biological material Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000007398 colorimetric assay Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- -1 halide ions Chemical class 0.000 description 3
- 229930195733 hydrocarbon Natural products 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000011664 signaling Effects 0.000 description 3
- 239000002356 single layer Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical group N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 150000007942 carboxylates Chemical class 0.000 description 2
- 150000004770 chalcogenides Chemical class 0.000 description 2
- 239000013626 chemical specie Substances 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 238000010339 medical test Methods 0.000 description 2
- CWQXQMHSOZUFJS-UHFFFAOYSA-N molybdenum disulfide Chemical compound S=[Mo]=S CWQXQMHSOZUFJS-UHFFFAOYSA-N 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000011045 prefiltration Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- 238000007829 radioisotope assay Methods 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 229910052582 BN Inorganic materials 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- PZNSFCLAULLKQX-UHFFFAOYSA-N Boron nitride Chemical compound N#B PZNSFCLAULLKQX-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 238000001327 Förster resonance energy transfer Methods 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 239000003131 biological toxin Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 150000005829 chemical entities Chemical class 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000009295 crossflow filtration Methods 0.000 description 1
- 239000002178 crystalline material Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000011229 interlayer Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000000670 ligand binding assay Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000002159 nanocrystal Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 150000001282 organosilanes Chemical class 0.000 description 1
- 239000011236 particulate material Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001846 repelling effect Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000009919 sequestration Effects 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003462 sulfoxides Chemical class 0.000 description 1
- 229910052717 sulfur Chemical group 0.000 description 1
- 239000011593 sulfur Chemical group 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
- G01N15/0272—Investigating particle size or size distribution with screening; with classification by filtering
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
- G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
Definitions
- the present disclosure generally relates to devices configured for withdrawal and/or dispensation of a fluid, particularly a medical fluid, and analysis thereof, and, more specifically, to syringes and other devices employing one or more two-dimensional separation membrane and methods for use thereof in separating and assaying for target entities of various sizes or chemical activities.
- Devices herein include those which can capture sub-micron materials, including nanosized materials from a fluid.
- Devices herein can function to selectively collect target entities within one or more predetermined range of sizes.
- predetermined size ranges may be representative of certain entity types, biological cells (protozoa, fungi, bacteria, mammalian cells, tumor cells), viruses (retrovirus, enveloped virus), biological molecules (e.g., proteins, polypeptides, nucleic acids, polysaccharides, peptide toxins), small molecules (e.g., drugs, chemical toxins), atomic species (e.g., halide ions, metal ions).
- Target entities collected by size range can be subjected to one or more assays appropriate for the type and size of entity collected.
- Target entities also referred to herein as analytes
- Target entities that are smaller than the occlusion size of a separation membrane
- Benefits to efficiency and selectivity in analysis can be obtained when target entities are separated by size range prior to assay, in that assays appropriate for target entities of a particular size range, e.g., biological cells, can be more selectively applied.
- the present disclosure describes filtration device configurations and methods for separating and assaying target entities having different sizes and/or chemical
- filtration device configurations include one or more filter membranes (also called separation membranes) disposed is series with one another where the filter membrane contain perforated two-dimensional material and wherein the filter membranes have an effective pore size that decreases in a directed of intended fluid flow.
- filter membranes also called separation membranes
- filtration device configurations include more than two filter membranes which function for size separation and which in combination separate entities in the fluid (including target entities) into one, or preferably more than one, size-range-selected pools of entities (including one or more target entities).
- the methods can include providing one or more filter membranes disposed in series with one another, where the filter membranes contain a perforated two-dimensional material and the filter membranes have an effective pore size that decreases in a direction of intended fluid flow; passing a fluid through the filter membranes; and optionally assaying for at least one target entity sequestered by the filter membranes. Assaying can take place while the at least one target entity is sequestered on the filter membranes or after it has been released therefrom. In a related embodiment, the sequestered at least one target entity can be selectively subjected to alteration which results in product entities thereof which product entities can be subject to subsequent size-separation and/or subject to one or more appropriate assays.
- more than two filtration membranes are disposed in series where effective pore size of a filter decreases in a direction of fluid flow where the filter membranes function in combination to separate or sequester a plurality of entities in the fluid into size-range- selected pools of entities (including one or more target entities).
- One or more assays can be applied to one or more of the size- selected pools of entities. Assays can be performed while the at least one target entity is sequestered on the filter membranes or after an entity has been released therefrom.
- the present disclosure also describes methods for administering a fluid to a patient.
- the methods can include providing at least one filter membrane containing a perforated two-dimensional material, and administering a fluid to a patient after passing the fluid through the at least one filter membrane, where the at least one filter membrane removes at least one biological material or toxin from the fluid.
- FIGURE 1A shows an illustrative schematic of a syringe containing a standard interface to which a filter membrane can be attached
- FIGURE IB shows an illustrative schematic of a syringe having a removable filter membrane containing a perforated two- dimensional material attached thereto;
- FIGURES 2A-2C show illustrative schematics of filter membranes containing a perforated two-dimensional material disposed between two layers of a support;
- FIGURES 3 and 3B show illustrative schematics of a graphene-based filter membrane disposed in a luer-lock housing
- FIGURE 4 shows an illustrative schematic of graphene-based filter membranes disposed in series, where the pore size can be the same or different;
- FIGURE 5 shows an illustrative schematic of a plurality of filter membranes arranged in series, where the effective pore size decreases in the direction of intended fluid flow;
- FIGURE 6 shows a schematic illustrating the effect of decreasing pore size, where progressively smaller molecular entities are occluded within the filter
- FIGURE 7 shows an illustrative schematic wherein the filter membrane configuration of FIGURE 5 can be stimulated by an electrical current to promote release and analysis of the target entities occluded therein;
- FIGURE 8 shows an illustrative schematic of a series of filter membranes stacked together to sequester biological entities of different effective sizes.
- the present disclosure is directed, in part, to devices containing one or more filter membranes containing a two-dimensional material.
- the present disclosure is also directed, in part, to methods for separating and optionally assaying target entities having a defined size or chemical characteristic from a fluid medium, particularly a biological fluid, using one or more filter membranes, where the filter membranes are each configured to separate target entities having a defined size or chemical characteristic.
- Graphene has garnered widespread interest for use in a number of applications due to its favorable mechanical and electronic properties.
- Graphene represents an atomically thin layer of carbon in which the carbon atoms reside as closely spaced atoms at regular lattice positions.
- the regular lattice positions can have a plurality of defects present therein, which can occur natively or be intentionally introduced to the graphene basal plane.
- defects will also be equivalently referred to herein as “apertures,” “perforations,” or “holes.
- the term “perforated graphene” will be used herein to denote a graphene sheet with defects in its basal plane, regardless of whether the defects are natively present or intentionally produced.
- graphene and other two-dimensional materials can represent an impermeable layer to many substances. Therefore, if they can be sized properly, the apertures in the impermeable layer can be useful retaining target entities that are larger than the effective pore size.
- a number of techniques have been developed for introducing a plurality of perforations in graphene and other two-dimensional materials, where the perforations have a desired size, number and chemistry about the perimeter of the perforations. Chemical modification of the apertures can allow target entities having particular chemical characteristics to be preferentially retained or rejected as well.
- the invention employs filtration membranes which comprise perforated two- dimensional materials with a plurality of apertures to effect separation of sub-micron or nano- sized components.
- Various two-dimensional materials useful in the present invention are known in the art.
- the two-dimensional material comprises graphene, molybdenum sulfide, or boron nitride.
- the two-dimensional material is a graphene-based material.
- the two-dimensional material is graphene.
- Graphene can include single-layer graphene, multi-layer graphene, or any combination thereof.
- nanomaterials having an extended two-dimensional molecular structure can also constitute the two-dimensional material in the various embodiments of the present disclosure.
- molybdenum sulfide is a representative chalcogenide having a two-dimensional molecular structure
- other various chalcogenides can constitute the two-dimensional material in the embodiments of the present disclosure.
- Choice of a suitable two-dimensional material for a particular application can be determined by a number of factors, including the chemical and physical environment into which the graphene or other two-dimensional material is to be terminally deployed.
- the two dimensional material useful in membranes herein is a sheet of graphene-based material.
- Graphene-based materials include, but are not limited to, single layer graphene, multilayer graphene or interconnected single or multilayer graphene domains and combinations thereof.
- graphene-based materials also include materials which have been formed by stacking single or multilayer graphene sheets.
- multilayer graphene includes 2 to 20 layers, 2 to 10 layers or 2 to 5 layers.
- graphene is the dominant material in a graphene-based material.
- a graphene-based material comprises at least 30% graphene, or at least 40% graphene, or at least 50% graphene, or at least 60% graphene, or at least 70% graphene, or at least 80% graphene, or at least 90% graphene, or at least 95% graphene.
- a graphene- based material comprises a range of graphene selected from 30% to 95%, or from 40% to 80% from 50% to 70%, from 60% to 95% or from 75% to 100%.
- a "domain" refers to a region of a material where atoms are uniformly ordered into a crystal lattice. A domain is uniform within its boundaries, but different from a neighboring region.
- a single crystalline material has a single domain of ordered atoms.
- at least some of the graphene domains are nanocrystals, having domain size from 1 to 100 nm or 10-100 nm.
- at least some of the graphene domains have a domain size greater than 100 nm to 1 micron, or from 200 nm to 800 nm, or from 300 nm to 500 nm. "Grain boundaries" formed by crystallographic defects at edges of each domain differentiate between neighboring crystal lattices.
- a first crystal lattice may be rotated relative to a second crystal lattice, by rotation about an axis perpendicular to the plane of a sheet, such that the two lattices differ in "crystal lattice orientation".
- the sheet of graphene-based material comprises a sheet of single or multilayer graphene or a combination thereof.
- the sheet of graphene-based material is a sheet of single or multilayer graphene or a combination thereof.
- the sheet of graphene-based material is a sheet comprising a plurality of interconnected single or multilayer graphene domains.
- the interconnected domains are covalently bonded together to form the sheet.
- the sheet is polycrystalline.
- the thickness of the sheet of graphene-based material is from
- a sheet of graphene-based material comprises intrinsic defects.
- Intrinsic defects are those resulting from preparation of the graphene-based material in contrast to perforations which are selectively introduced into a sheet of graphene-based material or a sheet of graphene.
- Such intrinsic defects include, but are not limited to, lattice anomalies, pores, tears, cracks or wrinkles.
- Lattice anomalies can include, but are not limited to, carbon rings with other than 6 members (e.g. 5, 7 or 9 membered rings), vacancies, interstitial defects (including incorporation of non-carbon atoms in the lattice), and grain boundaries.
- membrane or membrane portions comprising the sheet of graphene-based material further comprises non-graphenic carbon-based material located on the surface of the sheet of graphene-based material.
- the non-graphenic carbon-based material does not possess long range order and may be classified as amorphous.
- the non-graphenic carbon-based material further comprises elements other than carbon and/or hydrocarbons.
- Non-carbon elements which may be incorporated in the non-graphenic carbon include, but are not limited to, hydrogen, oxygen, silicon, copper and iron.
- the non-graphenic carbon-based material comprises hydrocarbons.
- carbon is the dominant material in non-graphenic carbon-based material.
- a non-graphenic carbon-based material comprises at least 30% carbon, or at least 40% carbon, or at least 50% carbon, or at least 60% carbon, or at least 70% carbon, or at least 80% carbon, or at least 90% carbon, or at least 95% carbon.
- a non- graphenic carbon-based material comprises a range of carbon selected from 30% to 95%, or from 40% to 80%, or from 50% to 70%.
- perforated Two-dimensional materials in which pores are intentionally created are referred to herein as "perforated”, such as “perforated graphene-based materials", “perforated two-dimensional materials' or “perforated graphene.”
- the present disclosure is also directed, in part, to perforated graphene, perforated graphene-based materials and other perforated two-dimensional materials containing a plurality of apertures (or holes) ranging from about 5 to about 1000 angstroms in size.
- the hole size ranges from lOOnm up to 1000 nm or from 100 nm to 500 nm.
- the present disclosure is further directed, in part, to perforated graphene, perforated graphene-based materials and other perforated two- dimensional materials containing a plurality of holes ranging from about 5 to 1000 angstrom in size and having a narrow size distribution, including but not limited to a 1-10% deviation in size or a 1-20% deviation in size.
- the characteristic dimension of the holes is from 5 to 1000 angstrom.
- the characteristic dimension is the diameter of the hole.
- the characteristic dimension can be taken as the largest distance spanning the hole, the smallest distance spanning the hole, the average of the largest and smallest distance spanning the hole, or an equivalent diameter based on the in-plane area of the pore.
- perforated graphene-based materials include materials in which non-carbon atoms have been incorporated at the edges of the pores.
- separation of various target entities can be desirable in a number of instances, particularly in biological separation processes.
- Such separation can be achieved employing a filter device having one or more or preferably more than two filter membranes disposed in series with one another, the filter membranes are spaced apart from each other, and the filter membranes having selected effective pore size that decreases in a direction of intended fluid flow wherein effective pore sizes of filters are selected to provide for separation of fluid components into pre-determined size-range pools.
- the filter membrane are spaced apart such that entities of a given size that pass through the pores of a preceding membrane or membranes are trapped or sequestered on a following membrane having pores sized such that the entities do not pass there through.
- each filter membrane comprises perforated two-dimensional material which functions for size selection.
- Spacing apart of the filter membrane can provide a space or enclosure for containment of entities sequestered on a filter after separation from fluid.
- This space or enclosure can be accessed generally for collection, analysis and/or identification of the sequestered entities, for example for collection of all or part of the sequestered entities, for introduction of light (of selected wavelength or wavelength range) to conduct an assay, for introduction of reagents or other materials for conducting an assay, or for observation of a change in color, wavelength of light introduced or other indicator associated with an assay.
- the space or enclosure can be accessed to modify one or more entities sequestered therein.
- Modification can include among others, reaction or interaction with a reagent or other added chemical or biological molecule, irradiation to break one or more bonds, release or braking of a bond by introduction of a reactant, introduction of light of selected wavelength, introduction of a ligand or antibody to bind to one or more entities sequestered.
- modification relates to application of an electrically current to one or more filter membranes.
- each filter membrane comprises perforated two dimensional material which is functionalized and wherein the filter membrane functions for separation by size and/or chemical characteristic.
- Functionalization includes functionalization in the vicinity of pores and/or functionalization on other portions of the filter membrane. Functionalization of filter pores can be accomplished by any means known in the art. Functionalization includes functionalization to attach carboxylate or related acidic or negatively charged chemical species or to attach amine or related basic or positively charged chemical species. Additional functionalization can include functionalization with hydrophobic groups or functionalization with hydrophilic groups where various such groups are known in the art. Additional functionalization can include functionalization with polar groups or functionalization with non-polar groups where various such groups are known in the art.
- Additional functionalization includes borate, sulfate, sulfoxide, and organosilanes among others.
- Functionalization can include functionalization with organic polymers or biological polymers.
- Functionalization includes functionalization to attach a protein receptor, a ligand, an antibody, or other chemical or biological species which selectively binds to one or more target entities.
- Functionalization is typically attached to the filter membrane or pores therein via a linking species which spaces the functionalization from the filter surface.
- Various linkers are known in the art and include hydrocarbon linkers, ether linkers, thioether linkers.
- a linker may contain a plurality of -CH 2 - moieties in combination with one or more -0-, -S-, -CO-, -COO-, -NH-, -NH-CO-.
- exemplary linkers can contain 2-50 carbon atoms and 2-20 heteroatoms selected from oxygen, nitrogen and sulfur.
- Exemplary useful size-range pools include (1) those that separate intact cells from the remains of disrupted cells (e.g., cell organelles, cell parts or cell components) or biological molecules contained in cells (e.g., nucleic acids, proteins, protein aggregates) or small molecules such as drugs or toxins; (2) those that separate different sizes of biological molecules (e.g., different size proteins, different size nucleic acids, different sizes of carbohydrates); (3) those that separate polymeric biological molecules (proteins, nucleic acids, polysaccharides) from non-polymeric biological molecules such as amino acids, small peptides, nucleotide, nucleosides, small nucleic acids (e.g., having 2-20 bases), monosaccharide, disaccharides or the like; (4) those that separate polymeric biological molecules from small molecules such as drugs or non-peptide toxins; or (5) those that separate protein receptors from ligands that potentially bind to such receptors. It will be apparent to one of ordinary skill in the art that many other size range pools may
- size-range pools include one or more pools containing entities ranging in size as follows: above 1000 nm; below 1000 nm, above 500 nm, below 500 nm., above 100 nm, below 100 nm, above 50 nm, below 50 nm, above 20 nm, below 20 nm, above 10 nm, below 10 nm, above 5 nm, below 5 nm, below 1 nm, between 1000 and 500 nm, between 500 and 100 nm, between 100 and 20 nm, between 20 nm and 10 nm, between 20 nm and 5 nm, between 5 nm and 1 nm, between 7-15 nm,.
- the range above 1000 nm or above 500 nm can be employed to capture biological cells.
- the range above 20 nm, the range between 100 and 20 nm or the range between 50 and 20 nm can be used to capture viruses.
- the range below 20 or between 4-20 nm can be used to capture proteins.
- Effective pores sizes of filter membranes can be selected to provide for separation into such exemplary size-range pools. It will be apparent to one of ordinary skill in the art that many other size range pools may be of interest and can be provides by appropriate choice of effective pore sizes.
- a filter device comprises a plurality of filter modules disposed in fluid communication and in series with one another along a direction of intended fluid flow wherein each filter module comprises a perforated two-dimensional material and a filter housing for holding the perforated two-dimensional material in place wherein the effective pore size of the perforated two dimension material of the serially disposed modules decrease in the direction of intended flow.
- the filter housings are configured for serial engaging or interfacing with adjacent filter housing to form a seal there between to prevent leakage of fluid when fluid is passaged through the filter device.
- each filter module is provided with an optionally valved inlet and an optionally valved outlet to facilitate fluid flow through the device.
- Valved inlets and outlets can be selectively opened or closed as desired. Closing of inlet and outlet valves in a module can be employed to isolate entities therein from those in other modules.
- the filter device includes a first and last filter module and intervening filter modules wherein the first module is provided with an optionally valved fluid inlet to facilitate fluid flow through the device and wherein the last module (with smallest pore size) is provided with an optionally valved fluid outlet.
- Inlets and outlets herein are optionally valved to allow for selective opening and closing thereof. Actuation of such valves can be by any known meaning and can be automated as known in the art and the opening and closing of selected valves can be optionally synchronized as known in the art. Inlets and outlets herein can optionally be provided as one-way valves, for example, to implement fluid flow (or predominant fluid flow) in one selected direction.
- At least one filter module of the filter device optionally further comprises an access port providing access to entities collected on the filter membrane.
- the access port is positioned such that it is not in the intended direction of fluid flow though the device.
- the access port opening is perpendicular to the intended direction of fluid flow through the device.
- the access port can be used for removal or addition to the filter module.
- the access port can be employed for removal of all or part of one or more target entities on the filter membrane.
- the access port can be employed for addition of light, particularly light of selected wavelength, e.g., UV-VIS, for example for modification or assay of one or more target entities.
- the access port can be employed for observing a color change, for collecting light and measuring wavelength and/or intensity, for collecting entitles sequestered on the filter membrane or products generated from entities.
- At least one filter module of the filter device further comprises a chamber formed adjacent to the filter membrane and in which entitles collected on the filter membrane can be enclosed.
- a chamber formed adjacent to the filter membrane and in which entitles collected on the filter membrane can be enclosed.
- Such chamber must provide for fluid flow through the device in the intended direction of fluid flow and as such is optionally provided with optionally valved fluid inlet and outlet.
- the chamber can however be formed between two adjacent filter modules which interface with each other via a connector or seal that prevents fluid leakage.
- a filter module can be provided with an optionally valved cross-flow inlet and/or an optionally valved cross-flow outlet.
- Such inlet or outlet can allow cross-flow of a fluid, such as a wash flow, other than the fluid from which target entities are to be separated or sequestered.
- Operation of a cross-flow outlet can be employed to selectively divert fluid flow from passage through subsequent filter modules disposed in the device. Coordinated operation of a cross-flow inlet and outlet can be used to flow a fluid through the folder and transverse to the filter membrane for example, to release entities including target entities from the surface of the membrane.
- the cross-flow can also be employed to introduce one or more selected reagents or reactants to the filter module to facilitate assay of the entities including target entities sequestered on the surface of the membrane.
- the flow emanating from the cross-flow outlet can be directed to a reservoir or other container for collection, disposal or additional processing as desired.
- the flow emanating from the cross-flow outlet for example, be directed into another filter module of the device or into a separate filtration device, or into an analytical instrument (i.e., Gas chromatograph (GC), mass spectrometer (MS) or GC/MS for analysis or further analysis.
- GC Gas chromatograph
- MS mass spectrometer
- GC/MS Gas chromatograph
- the fluid exiting the filtration device can be directed to a reservoir or other container for collection, disposal or further processing as desired or the exiting fluid can be directed to an analytical instrument for separation and/or analysis.
- the effective pore size of the filter membranes ranges in size from 0.5 nm to 1000 nm. In other specific embodiments, the effective pore size of the filter membranes range from 0.5 to 500 nm or from 0.5 to 100 nm or from 0.5 to 50 nm, or from 0.5 to 20 nm.
- a filter module of the filtration device is removable or replaceable with another filter module. In an embodiment, the number of filtration modules in the filtration device can be selectively changed by removal of one or more selected modules or by addition or one or more additional modules. Thus, the size-ranges that a given filter device can separate can be adjusted by addition or subtraction of one or more filter modules.
- a filtration device can be provided with a set of filter modules to provide a first set of pre-determined size-range separations and be modified by addition or subtraction of one or more filter modules for selected size ranges not in said first set to provide a second set of pre-determined size-range separations.
- the modular configuration of the filter device provide for significant flexibility in sculpting the size ranges that are to be separated in to size range pools.
- one or more assays are conducted in one or more filter modules of the filter device.
- one or more colorimetric assays are conducted in one or more filter modules of the invention.
- the housing of one or more filter modules in the filter device is transparent to allow observation of a color change associated with an assay.
- Various colorimetric assays (where a color change identifies a characteristic of one or more target entities) are known in the art an can be readily adapted for use in the devices and methods herein.
- the filter device of the invention is implemented in a syringe configuration.
- a luer lock fitting as an inlet for fluid flow in the intended direction can be employed.
- a fluid contained in a syringe barrel can be employed to provide fluid flow through the filter device.
- fluid can be drawn into the filter device by operation of the syringe plunger.
- fluid drawn into a syringe barrel can thereafter be pushed through the filter device by operation of the syringe plunger.
- a syringe equipped with a replaceable filter cartridge containing graphene can be used in separating target entities with a particular size range or certain chemical characteristics.
- Other perforated two-dimensional materials may be used in a similar manner.
- the filter cartridge can be interchangeable with filtration membranes of different perforation sizes and/or chemistries, such that a desired separation process can take place.
- a filter membrane with a first perforation size is inserted into the syringe and a fluid is drawn into the syringe body, such that target entities larger than the first perforation size are rejected on the membrane.
- a specimen of blood plasma can be withdrawn such that target entities smaller than the first perforation size are drawn into the syringe body.
- the filter membrane can be switched with a filter membrane having a second perforation size that is smaller than the first perforation size.
- the syringe rejects target entities on the filter membrane that are smaller than the second perforation size and molecular entities between the first perforation size and the second perforation size are dispensed for analysis.
- the fluid drawn into the syringe can be analyzed further without undergoing a subsequent separation process (e.g. , without being dispensed through the filter membrane having the second perforation size), although this approach has the potential for interference from smaller target entities that would have otherwise been removed with the second filter membrane.
- further separation of the fluid can take place before analyzing for a particular component.
- viruses can be separated from the dispensed fluid by conventional biological separation techniques, and the remaining components can then undergo analysis, such as specific quantitative or qualitative protein analyses. Any number of biological entities such as, for example, bacteria, viruses, protozoa, proteins, antibodies, and the like can be assayed through the techniques described herein.
- an assay is an investigative/analytical procedure in laboratory medicine, pharmacology, environmental biology and molecular biology for qualitatively assessing or quantitatively measuring the presence or amount, or the functional activity of a target entity, particularly a molecular entity.
- the target entity is sometimes referred to as an analyte or the measurand or the target of the assay.
- the target entity within a medium such as a fluid medium, often needs to be accumulated and separated from other fluid components such that the target entity can be further analyzed with sufficient detection sensitivity.
- the further analysis of the separated target entity can represent conventional medical assay techniques or analyses based upon nanotechnology.
- suitable filter membranes can be inserted into or attached to a syringe using a connection mechanism, which can allow filter membranes of various suitable sizes to be attached to the syringe.
- Suitable connection mechanisms will be familiar to one having ordinary skill in the art.
- suitable connection mechanisms can include a luer lock fitting.
- Other friction or compression fit connections can also be suitable for practicing the embodiments described herein, for example.
- one or more filter membranes containing graphene and/or another two-dimensional material can be stacked upon one another.
- connections between the filter membranes can be made via a standard fitting, such as a luer lock fitting on a housing in which the filter membrane is held.
- the filter membranes can include a single sheet of perforated graphene or other two- dimensional material, or multiple sheets (up to about 20 sheets). When multiple sheets are present, the perforation size (effective pore size) within each sheet can be the same or different, which can allow the interlayer flow to be altered. Moreover, the perforation size within each filter membrane can also the same or different as described hereinafter.
- the filter membranes can be disposed perpendicular to a fluid flow pathway (i.e., the fluid flow passes through a needle into the filter membranes and then into the syringe body).
- a fluid flow pathway i.e., the fluid flow passes through a needle into the filter membranes and then into the syringe body.
- cross-flow filtration configurations can be used. Cross-flow can also be used for purging or flushing of perpendicular disposed filter membranes.
- the graphene or other two-dimensional material can be functionalized.
- the perimeter of the apertures within the graphene can be functionalized.
- Suitable techniques for functionalizing graphene will be familiar to one having ordinary skill in the art.
- a skilled artisan will be able to choose a suitable functionality for producing a desired interaction with a target entity in a fluid, such as a biological fluid.
- the apertures in a graphene can be functionalized such that they interact preferentially with a protein or class of proteins in deference to other biological entities of similar size, thereby allowing separations based upon chemical characteristics to take place.
- a target entity can be captured within the filter membrane for further analysis, if desired, optionally by functionalizing the membrane material.
- the methods described herein can further include releasing the target entity or a signaling entity from the membrane material in order for analysis to take place. In other various embodiments, analysis can take place while the target entity is disposed on the filter membrane.
- the graphene or other two-dimensional material can be functionalized with a chemical entity so that the functionalization interacts preferentially with a particular type of biological target entity (e.g., by a chemical interaction).
- the graphene or other two-dimensional material can be functionalized such that it interacts electronically with a biological target entity (e.g., by a preferential electrostatic interaction).
- Graphene or other two-dimensional material may be treated with certain functionalization so as to repel/impede or attract/facilitate passage of components contained within a fluid, for example, allowing or facilitating certain components to pass through the membrane while repelling or impeding passage of undesired components.
- pores functionalized with negatively charged moieties such as carboxylate groups (-COO-) can repel or impede species that are positively charged (cationic).
- pores functionalized with positively charged species such as protonated amine groups, can repel or impede species that are negatively charged (anionic).
- the graphene or other two-dimensional material can be mounted on a porous substrate to provide among other benefits mechanical support.
- the porous substrate has pores large enough to allow passage of any entities that are intended to be separated in the filter device.
- a step of pre-filtration of the fluid that is to be passaged through the filter device is undertaken to remove large particulate material. It will be appreciated that pre-filtration should be selected to avoid removal of target entitles.
- a fluid that has been passaged through the filtration device may be subjected to one or more additional filtration step thereafter. The one or more additional filtration steps may be accomplished using a similar filtration device configuration or a different filtration device configuration.
- a fluid passaged through a filtration device of the invention maybe subsequently passaged through a sterilization filter (as are known in the art) to ensure elimination/exclusion of undesired microorganisms.
- the graphene or other two-dimensional material can be mounted on a substrate that facilitates detection, not just sequestration of a particular target entity.
- Suitable substrates for facilitating detection can encompass those providing for visible, colorimetric, fluorescent, UV-VIS or other confirmation and quantification of binding of specific target entities thereon, particularly those that have a size above the perforation size of the graphene.
- Activation of the assay for such detection mechanisms can take place by any number of factors such as, for example, time, temperature, electrical activation, and the like. For example, electrical power (e.g.
- a battery supplied by a battery
- Release of dye molecules for example, can be indicative of the presence or absence of a target entity on the graphene.
- filter membranes configured for detection of target entities of variable size or chemical characteristics can be stacked upon one other, where a flow pathway through the filter membranes progresses from the largest effective pore size to the smallest effective pore size.
- filter membranes configured for retaining and assaying a subject's blood for bacteria e.g. , e coli
- viruses e.g. , hepatitis or HIV
- radioisotopes or heavy metals can be disposed in series with one another, as depicted in FIGURE 8.
- Other biological entities such as antibodies and the like can also be separated and analyzed in a similar manner.
- a fluid containing target entities to be analyzed can be drawn into a syringe to which is attached a plurality of filter membranes, where an effective pore size of the filter membranes progresses from largest to smallest.
- a needle can be attached to the filter membrane having the largest effective pore size and the syringe can be attached to the filter membrane having the smallest effective pore size.
- Target entities trapped within the filter membranes can then undergo further analysis, as described hereinafter, or the fluid in the syringe can be assayed.
- the target entities trapped therein can be analyzed individually, thereby decreasing the opportunity for analytical interference.
- a fluid containing target entities to be analyzed can be drawn into a syringe or like fluid withdrawal device without first being filtered. Thereafter, a plurality of filter membranes can be attached to the syringe, where an effective pore size of the filter membranes progresses from largest to smallest and the filter membrane with the largest effective pore size is attached to the syringe.
- the filter membranes can be initially connected to the syringe and connected to one another in the same order, but the filter membranes can be bypassed when initially drawing the fluid into the syringe. In either case, the fluid in the syringe can be passed through the filter membranes starting with the largest effective pore size and proceeding to the smallest effective pore size. As before, the trapped target entities can then undergo further analysis.
- the target entities can then be assayed according to the embodiments described herein.
- Assays can take place using common assay techniques that will be familiar to one having ordinary skill in the art, such as assays common in the scientific literature and routinely practiced in the laboratory.
- the assay can be activated by time, temperature, electrical or other activation mechanism, as further described above.
- this feature can result in fixing/electrically immobilizing the graphene or the substrate to result in separation of the various sections. Assays of the separated target entities can then be conducted through any suitable analysis technique, including those based upon nanotechnology.
- kits that represents a modular test that can be disposable and provide easy to read results, with the opportunity for various levels of customization and modification to suit a particular testing application.
- the filter membranes described herein can be used to further improve patient safety.
- the filter membranes can be used to remove viruses, bacteria or other pathogens from a fluid being administered to a patient, so as to prevent the spread of disease.
- a syringe can be used for administering a fluid to a patient, it is to be recognized that other dispensation devices can also be used similarly, such as IV bags, infusion pumps, and the like.
- FIG. 1A shows an illustrative schematic of a syringe (10) containing a standard interface to which a filter membrane can be attached for example, within a filter module (30).
- the syringe includes a needle (20) shown as attached to the syringe.
- FIG. IB shows an illustrative schematic of a syringe having a removable filter membrane containing a perforated two-dimensional material attached thereto, as illustrated within a filter module (30).
- a syringe useful in the present invention is designated generally numeral 10.
- the syringe (10) has a barrel (12) which is of a tubular construction.
- the barrel (12) has a plunger end (14) that is opposite a needle end (16).
- the barrel (12) provides an open interior 18. Extending radially from the plunger end (14) is a flange (19) to facilitate manual operation of the plunger (24).
- Hub (17) provides for connection to the needle (20) which connection can be made by various standard connection interfaces (21), including a luer lock connection.
- the plunger (24) is slidably received in the barrel (12).
- the plunger (24) includes a plunger tip (26) at one end which has an outer diameter sized to allow slidable movement within the interior (18).
- the plunger tip (26) is sized to create enough of a seal to preclude migration of material from within the interior (18) while also generating a suction force at the needle end (16) when the plunger is pulled.
- a push end Opposite the plunger tip (26) is a push end (28).
- the push end (28) may be manipulated by a user, or an automated mechanism or the like to move the plunger in a desired direction.
- Suction mechanisms other than a plunger within a barrel may be utilized to pull or draw material through filter modules or filter membranes with pores as disclosed herein.
- One or more filter modules (30) is maintained at the needle end (16) of the barrel(l). Hub 17 is connected to an end of the filter cartridge 30 opposite the needle end 16 of the barrel.
- the filter module is provides with connector 31 (which can be any standard interface that is fluid tight, such as a luer lock fitting. Filtration is accomplished in a syringe device as shown by operation of the plunger to draw fluid into a needle, through the filter module and into the barrel of the syringe. Alternatively, a fluid can be first drawn into the syringe barrel prior to attachment of the filter module, then the filter module can be attached and fluid cam be pushed through the filter module.
- the direction of fluid flow in these alternative modes of operation is in opposite directions so that the order of a plurality of filter modules for size-range separations is appropriately adjusted.
- the filter membranes (and the filter modules containing them) are arranged in series with filter membrane pore size decreasing in the direction of fluid flow.
- the filter module (30) may be moveable and/or replaceable so as to allow for retention of desired size components, such as molecules, or a size range of components, such as molecules.
- the syringe functions for fluid flow control through a filter module, filter modules in series or a filter device.
- the one or more filter modules of this invention can be implemented in with a variety of filter flow control devices, for example one or more pumps with optional flow controls and appropriate fluid conduits can be provided to implement fluid flow control.
- FIGURES 2A-2C show illustrative schematics of filter membranes (40-A-C) containing a perforated two-dimensional material (45A-C) disposed between two layers of a porous support (41A-C and 42 A-C). While illustrated in certain shapes, the filter membrane can be any shape. The direction of flow is shown as 43 A. Note that a single layer of support preferably (layer 41A-41C) may be employed.
- the filter membrane in FIGURE 2A has a two-dimensional material 45A which is perforated to have apertures or pores 46.
- the filter membrane in FIGURE 2B has a two-dimensional material 45B which is perforated to have apertures or pores 47.
- the pores sizes of filter membranes in different modules are typically different and are selected to be different as discussed herein above.
- pores 46 have effective pore size larger than the effective pore size of pores 47.
- FIGURE 2C illustrates a specific embodiment of a filter membrane useful in the invention the use of which has been described in International application PCT/US2015/18114, filed February 27, 2015, which in turn claims the benefit of US application 14/193,007, filed February 28, 2014.
- the filter membrane 45 C in this illustrated embodiment contains two portions: a portion where the pores 46 are larger than the pores 47 in the second portion.
- Such a filter membrane can be used in a filter module where in the filter membrane is mounted in a holder having a mechanical mechanism for switching the two portions of the filter membrane in and out of the fluid flow.
- Various such mechanisms are shown in the patent documents cited above.
- FIGURES 3A and 3B show illustrative schematics of a perforated graphene- based filter membrane (40) disposed in a luer-lock housing (32) having luer lock fitting (33).
- the luer lock fitting provides an exemplary inlet.
- the filter membrane (40) is shown as supported on a porous support layer (41).
- the module has a chamber 49 which encloses entitles sequestered on the filter membrane after filtration.
- FIGURE 3B illustrates alternative filter module having a side port (55) which can be used to access the chamber with the module.
- This side port can function as an optionally valved inlet or outlet and as such can also be used for introducing other components, such as buffers, or for cross-flow purging, for example.
- a plurality of such side ports can be provided in a given filter modules.
- FIGURE 4 shows an illustrative schematic of graphene-based filter modules
- the pore size of the modules in series decreases in the direction of flow through the filter modules.
- pores size A is larger than pores B is larger than pore size C.
- the plurality of filter modules shown can be implemented in a filter device wherein the modules are interfaced one with the other via a fluid tight sealing mechanism.
- Such a filter device can further be provided with any of various optionally valves inlets and outlets to facilitate fluid flow through the filter.
- a plurality of filter modules can be implemented in a filter device employing a syringe to provide for fluid flow.
- FIGURE 5 shows an illustrative schematic of a plurality of filter membranes (30/50) arranged in series with a first and a last module and intervening modules (30/50A-E), where the effective pore size decreases in the direction of intended fluid flow.
- the first module can be provided with an optionally valved inlet (56) and the last module in series can be provided with an optionally valved outlet (57).
- This configuration provides a filter device 70.
- FIGURE 6 shows a schematic illustrating the effect of decreasing pore size, where progressively smaller molecular entities are occluded within the filter.
- binding of the target entities to the filter membrane can result in release of a dye or like signaling entity that can be detected to indicate the presence, absence or saturation of the target entity on the filter membrane.
- the dye can be released during a further stimulation event of the filter membrane, as discussed hereinafter and elsewhere herein.
- the occlusion of target entities by the filter membrane can permit Forster resonance energy transfer analyses to be conducted. As one of ordinary skill in the art will recognize, such measurements are based upon the distance between a fluorescent molecule and a quencher. Thus, by measuring fluorescence, the presence or absence of a target entity can be determined. Other suitable analysis techniques for assaying for the target entities on the filter membrane can also be envisioned.
- FIGURE 7 shows an illustrative schematic wherein the filter membrane configuration of FIGURE 5 can be stimulated by an electrical current to promote release and/or analysis of the target entities housed therein.
- Optional spacers 60 and 61 can be used to facilitate connection of a battery (62) to the filter device 70 which is composed of a plurality of filter modules (30/50).
- electrical stimulation is but one technique whereby further assay of the target entities may take place.
- Other sources of electrical power can be envisioned, such as a direct electrical connection to an AC or DC power source, a generator, a hand cranked generator, or the like.
- application of voltage can result in cell lysing to release molecules, modify the filter membranes to release the target entities or a dye/visualization molecule, or fix/immobilize/seal the graphene to make the filter membranes separable from one another.
- the simultaneous detection and analyses offered by the configurations of FIGURES 5 and 7 are believed to represent particular advantages in analyzing the complex mixtures that can often be present in biological media.
- FIGURE 8 shows an illustrative schematic of a series of filter modules stacked together forming a filter device 80.
- the filter module configuration is illustrated to sequester biological entities of different effective sizes and filter membrane pore size decreases in the direction of flow.
- Module 75A sequesters biological cells such as Escherichia coli (alternatively mammalian cells, including tumor cells might be sequestered).
- Modules 75B- 75 C is illustrated to sequester varying sizes of viruses/or biological polymers such as proteins
- Module 75E is illustrated to sequester small molecules, atoms or ions, such as heavy metal atoms or ions.
- FIGURE 8 also illustrates that the type of assay that may be performed can be selected as appropriate for the size and type of species sequestered in a given module.
- the assays noted in FIGURE 8 are illustrative.
- cells sequestered in module 75A can be removed from the module and identified by standard methods.
- cell captures in module 75A can be lysed (chemically or via application of electrical current) to collect nucleic acids or other biological polymers which can be assayed by well-known methods.
- hybridization assays or PCR (polymerase chain reaction) methods can be employed to identify the cells that have been captured. It is noted that an intermediate step of culturing of the cells capture may be applied to facilitate identification of captured cells.
- cell can be captures on a first filter module and the captures cells lysed, for example by application of electric current to the filter membrane.
- the lysate of the cells can then be flushed with application of a carrier fluid to another filter modules or series of filter modules to perform size-range separation on the cell lysate.
- Various known assays can be performed on size-range selected lysate components, such as radioisotope assays, chemical assays, ligand binding assays and the like.
- HIV test Various assays for the presence of viruses can be applied if desired (HIV test,
- Hep C test Various tests for the presence of a selected protein can be applied, for example, selective antibody assays for a given protein.
- a radioisotope assay may be employed on fluid samples which contain components which are enriched in such isotopes or where one or more selected components are treated to contain certain radioisotopes.
- a series of colorimetric assay are performed on the entities captures in the different modules and the results of any color change can be observed by observing a color change in the module or on a given filter membrane.
- the housings of the filter modules are transparent to allow observation of such color change.
- one or more assays that effect a change in fluorescence are employed and the change in fluorescence in the module or on the filter membrane can be observed or detected by methods that are known in the art.
- the filter device is configured such that entities capture within the module on the filter membrane of released form the filter membrane can be assayed by irradiation with light, e.g., UV-VIS spectroscopy.
- the filter device of the invention can be implemented in a kit providing a plurality of filter modules, with filters of selected sized for construction of a filter device for separation of selected size-ranges of entities.
- a kit can provide a set of filter modules having pore sizes A1-A20 , where Al is the largest pore size and A20 is the smallest pore size.
- the pore sizes of the filter modules can be selected to cover a desired range of sizes, for example from one to 1000 nm and the pore sizes of modules can be set at intermediate pore sizes in this range (e.g., A20 at 50 nm, A19 at 100 nm, A18 at 150 nm...Al at 1000 nm) and two or more of the filter modules can be arranged in decreasing size order to form a selected filter device to provide desired size range pools (e.g., A20, A15, A10, A5, Al in series; or A18, A7, A5, A2, Al in series.) It will be appreciated that various combinations of filter modules can be combined to achieved desired size-range separations.
- FIGURE 7 illustrates application of a current to all of the filter modules. However, it will be appreciated that current can be applied to fewer than all filter modules. In this embodiment, it will be appreciated that filter modules to which current is to be applied must be electrically isolated from those filter modules to which current is not to be applied. Appropriate isolation methods and materials to achieve electrical isolation arte known in the art. When a conductive two-dimensional material is employed in the filter membrane, such as a graphene- based material or graphene, then electrical leads can be provided as appropriate to the filter membrane itself to supply current thereto.
- a conductive two-dimensional material is employed in the filter membrane, such as a graphene- based material or graphene
Abstract
Description
Claims
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA2947029A CA2947029A1 (en) | 2014-05-01 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
AU2015252784A AU2015252784A1 (en) | 2014-05-01 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
EP15786691.4A EP3137188A4 (en) | 2014-05-01 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
JP2016565216A JP2017519974A (en) | 2014-05-01 | 2015-05-01 | Separation and analysis of target substances using filter membranes with perforated two-dimensional materials |
CN201580021974.0A CN106457078A (en) | 2014-05-01 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
US15/308,351 US20170067807A1 (en) | 2014-02-28 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
US15/099,588 US10653824B2 (en) | 2012-05-25 | 2016-04-14 | Two-dimensional materials and uses thereof |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201461987410P | 2014-05-01 | 2014-05-01 | |
US61/987,410 | 2014-05-01 | ||
USPCT/US2015/018114 | 2015-02-27 | ||
PCT/US2015/018114 WO2015131109A1 (en) | 2014-02-28 | 2015-02-27 | Syringe for obtaining sub-micron materials for selective assays and related methods of use |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2015/018114 Continuation WO2015131109A1 (en) | 2012-05-25 | 2015-02-27 | Syringe for obtaining sub-micron materials for selective assays and related methods of use |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US15/099,588 Continuation-In-Part US10653824B2 (en) | 2012-05-25 | 2016-04-14 | Two-dimensional materials and uses thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2015168653A1 true WO2015168653A1 (en) | 2015-11-05 |
Family
ID=54359410
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2015/028948 WO2015168653A1 (en) | 2012-05-25 | 2015-05-01 | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP3137188A4 (en) |
JP (1) | JP2017519974A (en) |
CN (1) | CN106457078A (en) |
AU (1) | AU2015252784A1 (en) |
CA (1) | CA2947029A1 (en) |
WO (1) | WO2015168653A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10005038B2 (en) | 2014-09-02 | 2018-06-26 | Lockheed Martin Corporation | Hemodialysis and hemofiltration membranes based upon a two-dimensional membrane material and methods employing same |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6699684B2 (en) * | 2002-07-23 | 2004-03-02 | Nalco Company | Method of monitoring biofouling in membrane separation systems |
US20060166347A1 (en) | 2005-01-27 | 2006-07-27 | Applera Corporation | Sample preparation devices and methods |
US7306768B2 (en) * | 2003-02-21 | 2007-12-11 | Filtertek Inc. | Filter for medical and laboratory use, especially for blood analysis and the like |
US20080045877A1 (en) * | 2006-08-15 | 2008-02-21 | G&L Consulting, Llc | Blood exchange dialysis method and apparatus |
US20110100921A1 (en) * | 2008-05-17 | 2011-05-05 | Hans-Werner Heinrich | Device for Separating Particles in and from Liquids and Use of Said Device in Biotechnology, Biological Research, Diagnostics and the Treatment of Diseases |
US8329476B2 (en) * | 2009-05-22 | 2012-12-11 | Teknologian Tutkimuskeskus Vtt | Sample port, multi-layer filter, sampling method, and use of a sample port in sampling |
US20130240355A1 (en) * | 2012-03-16 | 2013-09-19 | Lockheed Martin Corporation | Functionalization of graphene holes for deionization |
WO2013138698A1 (en) | 2012-03-15 | 2013-09-19 | Massachusetts Institute Of Technology | Graphene based filter |
US20130256211A1 (en) | 2012-03-29 | 2013-10-03 | Lockheed Martin Corporation | Tunable layered membrane configuration for filtration and selective isolation and recovery devices |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006262891A (en) * | 2004-11-30 | 2006-10-05 | D M L:Kk | Kit, method and device for assaying microorganism in liquid sample |
-
2015
- 2015-05-01 WO PCT/US2015/028948 patent/WO2015168653A1/en active Application Filing
- 2015-05-01 EP EP15786691.4A patent/EP3137188A4/en not_active Withdrawn
- 2015-05-01 CN CN201580021974.0A patent/CN106457078A/en active Pending
- 2015-05-01 AU AU2015252784A patent/AU2015252784A1/en not_active Abandoned
- 2015-05-01 CA CA2947029A patent/CA2947029A1/en not_active Abandoned
- 2015-05-01 JP JP2016565216A patent/JP2017519974A/en active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6699684B2 (en) * | 2002-07-23 | 2004-03-02 | Nalco Company | Method of monitoring biofouling in membrane separation systems |
US7306768B2 (en) * | 2003-02-21 | 2007-12-11 | Filtertek Inc. | Filter for medical and laboratory use, especially for blood analysis and the like |
US20060166347A1 (en) | 2005-01-27 | 2006-07-27 | Applera Corporation | Sample preparation devices and methods |
US20080045877A1 (en) * | 2006-08-15 | 2008-02-21 | G&L Consulting, Llc | Blood exchange dialysis method and apparatus |
US20110100921A1 (en) * | 2008-05-17 | 2011-05-05 | Hans-Werner Heinrich | Device for Separating Particles in and from Liquids and Use of Said Device in Biotechnology, Biological Research, Diagnostics and the Treatment of Diseases |
US8329476B2 (en) * | 2009-05-22 | 2012-12-11 | Teknologian Tutkimuskeskus Vtt | Sample port, multi-layer filter, sampling method, and use of a sample port in sampling |
WO2013138698A1 (en) | 2012-03-15 | 2013-09-19 | Massachusetts Institute Of Technology | Graphene based filter |
US20130240355A1 (en) * | 2012-03-16 | 2013-09-19 | Lockheed Martin Corporation | Functionalization of graphene holes for deionization |
US20130256211A1 (en) | 2012-03-29 | 2013-10-03 | Lockheed Martin Corporation | Tunable layered membrane configuration for filtration and selective isolation and recovery devices |
Non-Patent Citations (1)
Title |
---|
See also references of EP3137188A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10005038B2 (en) | 2014-09-02 | 2018-06-26 | Lockheed Martin Corporation | Hemodialysis and hemofiltration membranes based upon a two-dimensional membrane material and methods employing same |
Also Published As
Publication number | Publication date |
---|---|
AU2015252784A1 (en) | 2016-11-17 |
CN106457078A (en) | 2017-02-22 |
EP3137188A4 (en) | 2018-01-03 |
JP2017519974A (en) | 2017-07-20 |
EP3137188A1 (en) | 2017-03-08 |
CA2947029A1 (en) | 2015-11-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20170067807A1 (en) | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material | |
US20210291184A1 (en) | Apparatus, system and method for performing automated centrifugal separation | |
CN203030295U (en) | Test tube for mixing and extracting liquid | |
EP3350569B1 (en) | Flow cells utilizing surface-attached structures, and related systems and methods | |
CA2941101A1 (en) | Syringe for obtaining sub-micron materials for selective assays and related methods of use | |
AU2004267536B2 (en) | Apparatus for processing a fluid sample | |
JP2012504956A (en) | Cell sorting device | |
WO2016061453A1 (en) | Devices and methods for target analyte detection in liquid samples | |
WO2013158045A1 (en) | Microfilter and apparatus for separating a biological entity from a sample volume | |
AU703502B2 (en) | Method and apparatus for preparing substances for optical analysis | |
EP3137188A1 (en) | Separation and assay of target entities using filtration membranes comprising a perforated two-dimensional material | |
US11927600B2 (en) | Fluidic bridge device and sample processing methods | |
EP2720798B1 (en) | Processing of biological sample components | |
EP3793716A1 (en) | Systems and methods for nucleic acid purification using flow cells with actuated surface-attached structures | |
CN110506200A (en) | Method for filtering small size heterogeneous suspension in digital microcurrent-controlled device | |
CN109844491A (en) | Specimen separation device based on paper folding | |
WO2012065075A2 (en) | Electrokinetic devices and methods for high conductance and high voltage dielectrophoresis (dep) | |
WO2022051364A1 (en) | Label-free, real-time, whole-cell response monitoring with liquid raman spectroscopy | |
KR20170028443A (en) | Pretreatment method and nucleic acid extraction kit used in said method |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 15786691 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2947029 Country of ref document: CA |
|
ENP | Entry into the national phase |
Ref document number: 2016565216 Country of ref document: JP Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2015786691 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2015786691 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 15308351 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2015252784 Country of ref document: AU Date of ref document: 20150501 Kind code of ref document: A |