Classifications

• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
  • B01D15/08—Selective adsorption, e.g. chromatography
  • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
  • B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 and B01D15/30 - B01D15/36, e.g. affinity, ligand exchange or chiral chromatography
  • B01D15/3804—Affinity chromatography
• • 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
  • G01N1/405—Concentrating samples by adsorption or absorption
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/30—Processes for preparing, regenerating, or reactivating
  • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
  • B01J20/3291—Characterised by the shape of the carrier, the coating or the obtained coated product
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
  • B01J20/28002—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their physical properties
  • B01J20/28004—Sorbent size or size distribution, e.g. particle size
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
  • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
  • B01J20/28016—Particle form
  • B01J20/28021—Hollow particles, e.g. hollow spheres, microspheres or cenospheres
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
  • B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
  • B01J20/289—Phases chemically bonded to a substrate, e.g. to silica or to polymers bonded via a spacer
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/30—Processes for preparing, regenerating, or reactivating
  • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
  • B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
  • B01J20/3204—Inorganic carriers, supports or substrates
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/30—Processes for preparing, regenerating, or reactivating
  • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
  • B01J20/3214—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the method for obtaining this coating or impregnating
  • B01J20/3217—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
  • B01J20/3219—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond involving a particular spacer or linking group, e.g. for attaching an active group
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/30—Processes for preparing, regenerating, or reactivating
  • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
  • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
  • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
  • B01J20/3244—Non-macromolecular compounds
  • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
  • B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
  • B01J20/30—Processes for preparing, regenerating, or reactivating
  • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
  • B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
  • B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
  • B01J20/3244—Non-macromolecular compounds
  • B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
  • B01J20/3257—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one of the heteroatoms nitrogen, oxygen or sulfur together with at least one silicon atom, these atoms not being part of the carrier as such
  • B01J20/3261—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one of the heteroatoms nitrogen, oxygen or sulfur together with at least one silicon atom, these atoms not being part of the carrier as such comprising a cyclic structure not containing any of the heteroatoms nitrogen, oxygen or sulfur, e.g. aromatic structures
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
  • C07K1/14—Extraction; Separation; Purification
  • C07K1/16—Extraction; Separation; Purification by chromatography
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
  • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
  • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
  • C12N15/09—Recombinant DNA-technology
  • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
  • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
  • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
  • C12N15/101—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
  • B01J2220/00—Aspects relating to sorbent materials
  • B01J2220/80—Aspects related to sorbents specially adapted for preparative, analytical or investigative chromatography
  • B01J2220/82—Shaped bodies, e.g. monoliths, plugs, tubes, continuous beds

Definitions

• the present disclosure relates to compositions and methods useful for separating biomaterial from a sample.
• a composition for use in bioseparation comprises a plurality of hollow particles having a siliceous surface.
• the composition further comprises a surface-modifying agent bonded to the hollow particles.
• the surface-modifying agent comprises a binding segment and a reactive segment.
• the binding segment comprises a silyl group and the reactive segment comprises a reactive nitrogen group.
• the plurality of hollow particles have an average density between about 0.05 and 0.4 g/ml.
• a method for making a bioseparation composition comprises size fractionating a first plurality of hollow particles having a first particle size distribution with a first span to form a second plurality of hollow particles having a second particle size distribution with a second span that is less than the first span.
• the method further comprises surface-modifying either or both of the first and second plurality of hollow particles with a surface modifying agent to form surface-modified hollow particles.
• the surface-modifying agent comprises a binding segment and a reactive segment.
• the binding segment comprises a silyl group and the reactive segment comprises a reactive nitrogen group.
• a method for capturing an analyte comprises providing a bioseparation composition.
• the bioseparation composition comprises a plurality of hollow particles having a siliceous surface.
• the composition further comprises a surface-modifying agent bonded to the hollow particles.
• the surface-modifying agent comprises a binding segment and a reactive segment.
• the binding segment comprises a silyl group and the reactive segment comprises a reactive nitrogen group.
• the plurality of hollow particles have an average density between about 0.05 and 0.4 g/ml.
• the method further comprises contacting the bioseparation composition with a solution comprising an analyte.
• Typical isolation processes involve the introduction of particles into a sample (e.g., a solution) containing a biomaterial target, agitation of the sample, attachment of the biomaterial target to the particles, and subsequent separation of the particles from the sample. Separation of the particles, using known particle compositions, however, is time consuming and/or requires additional processing steps (e.g., centrifugation, filtration).
• bubble refers to a small, hollow globule, for example, a small spherical volume of gas encased within a thin film.
• analyte refers to any substance which may be present in a sample, and that it is desirable to separate from the sample or to detect in an assay.
• the analyte can be, without limitation, any substance.
• an analyte may comprise a substance for which there exists a naturally occurring antibody or for which an antibody can be prepared.
• the analyte may, for example, be a protein, a polypeptide, a hapten, a carbohydrate, a lipid, a drug, a bacterium, a virus, an enzyme, a cell, a cellular subcomponent or organelle (e.g., lysozomes, mitochondria) or any other of a wide variety of biological or non-biological molecules, complexes or combinations thereof.
• the analyte is a nucleic acid (DNA, RNA, PNA and nucleic acids that are mixtures thereof or that include nucleotide derivatives or analogs).
• alkyl refers to a monovalent group that is a radical of an alkane and includes groups that are linear, branched, cyclic, bicyclic, or a combination thereof.
• the alkyl group typically has 1 to 30 carbon atoms.
• the alkyl group contains 1 to 20 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms.
• the alkyl group contains one or more heteroatoms, such as oxygen, nitrogen, or sulfur atoms.
• alkylene refers to a divalent group that is a radical of an alkane and includes groups that are linear, branched, cyclic, bicyclic, or a combination thereof.
• the alkylene group typically has 1 to 30 carbon atoms.
• the alkylene group has 1 to 20 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms.
• the alkylene group contains one or more heteroatoms, such as oxygen, nitrogen, or sulfur atoms.
• alkyleneoxy refers to a divalent group that is an oxy group bonded directly to an alkylene group.
• alkoxy refers to a monovalent group having an oxy group bonded directly to an alkyl group.
• aryl refers to a monovalent group that is aromatic or heteroaromatic.
• the aryl has at least one unsaturated carbocylic or heterocyclic ring and can have one or more additional fused rings that can be unsaturated, partially saturated, or saturated.
• Aryl groups often have 6 to 20 carbon atoms, 6 to 18 carbon atoms, 6 to 16 carbon atoms, 6 to 12 carbon atoms, or 6 to 10 carbon atoms, and 0 to 5 heteroatoms selected from oxygen, sulfur, or nitrogen.
• arylene refers to a divalent group that is aromatic or heteroaromatic.
• the arylene has at least one unsaturated carbocylic or heterocyclic ring and can have one or more additional fused rings that can be unsaturated, partially saturated, or saturated.
• Arylene groups often have 6 to 20 carbon atoms, 6 to 18 carbon atoms, 6 to 16 carbon atoms, 6 to 12 carbon atoms, or 6 to 10 carbon atoms, and 0 to 5 heteroatoms selected from oxygen, sulfur, or nitrogen.
• aryloxy refers to a monovalent group having an oxy group bonded directly to an aryl group.
• aralkyl refers to a monovalent group that is an alkyl group substituted with an aryl group.
• Aralkyl groups often have an alkyl portion with 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms and an aryl portion with 6 to 20 carbon atoms, 6 to 18 carbon atoms, 6 to 16 carbon atoms, 6 to 12 carbon atoms, or 6 to 10 carbon atoms.
• aralkyloxy refers to a monovalent group having an oxy group bonded directly to an aralkyl group. Equivalently, it can be considered to be an alkoxy group substituted with an aryl group.
• aralkylene refers to a divalent group that is an alkylene group substituted with an aryl group or an alkylene group attached to an arylene group.
• Aralkylene groups often have an alkylene portion with 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms and an aryl or arylene portion with 6 to 20 carbon atoms, 6 to 18 carbon atoms, 6 to 16 carbon atoms, 6 to 12 carbon atoms, or 6 to 10 carbon atoms.
• acyloxy refers to a monovalent group of formula —O(CO)R b where R b is alkyl, aryl, or aralkyl.
• Suitable alkyl R b groups often have 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms.
• Suitable aryl R b groups often have 2 to 12 carbon atoms and 0 to 3 heteroatoms, such as, for example, phenyl, furyl, or imidazolyl.
• Suitable aralkyl R b groups often have an alkyl group with 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms that is substituted with an aryl having 6 to 12 carbon atoms such as, for example, phenyl.
• hydrolyzable group refers to a group that can react with water having a pH of 1 to 10 under conditions of atmospheric pressure.
• the hydrolyzable group is often converted to a hydroxyl group when it reacts.
• the hydroxyl group often undergoes further reactions.
• Typical hydrolyzable groups include, but are not limited to, alkoxy, aryloxy, aralkyloxy, acyloxy, or halo.
• the term is often used in reference to one of more groups bonded to a silicon atom in a silyl group.
• non-hydrolyzable group refers to a group that cannot react with water having a pH of 1 to 10 under conditions of atmospheric pressure.
• Typical non-hydrolyzable groups include, but are not limited to alkyl, aryl, and aralkyl.
• the present disclosure relates to compositions for use in bioseparation (e.g., isolating and/or assaying analytes).
• the compositions may include particles and one or more surface-modifying agents bonded to the particles.
• particles useful in the compositions of the present disclosure may include hollow particles having an at least partially solid outer region (e.g., shell) and a hollow inner region (e.g., core).
• useful particles may include bubbles having a substantially spherical hollow inner region encased by an outer region.
• the hollow inner region of the bubbles may be void of fluid, or be filled with a gas, including, but not limited to oxygen, nitrogen, carbon dioxide, helium, fluorocarbon gases and various combinations thereof, such as air.
• the outer region may be any material that can encase a volume of fluid, for example, a solid such as a metal, glass, ceramic, or similar material.
• the outer region may include a siliceous material having a siliceous surface (e.g., for bonding to a surface-modifying agent).
• the hollow particles may include glass bubbles, such as those sold by 3M under the trade designation SCOTCHLITE Glass Bubbles.
• the particles of the present disclosure may be configured and/or sized to facilitate rapid separation from a solution (e.g., based on buoyancy forces).
• the particles may have an average density of less than about 1 g/ml, less than about 0.8 g/ml, less than about 0.6 g/ml, or even less than about 0.4 g/ml.
• the particles may have an average density in a range of from about 0.05 g/ml to about 0.8 g/ml, or from about 0.08 g/ml to about 0.4 g/ml.
• the particles may have a mean particle size of less than about 200 micrometers, less than about 100 micrometers, or even less than about 80 micrometers.
• the particles may have a mean particle size in a range of from about 5 to 250 micrometers, from about 10 to 100 micrometers, or from about 20 to 80 micrometers.
• the particles useful in the compositions of the present disclosure may be a collection of particles that have been size fractionated (i.e., isolated based on size) from a larger collection of particles to achieve a desired particle size distribution.
• the width of a particle size distribution can be expressed by the following formula:
• 90P is the size for which 90 percent of the particles in the distribution are smaller (referred to as the 90th percentile size); 10P is the size for which only 10 percent of the particles in the distribution are smaller (referred to as the 10th percentile size); 50P is the size for which 50 percent of the particles in the distribution are smaller (referred to as the 50th percentile size); and GQ stands for the gradation quotient.
• the gradation quotient is also commonly known in the art by the term “span”.
• the particles useful in the bioseparation compositions of the present disclosure may have a size distribution with a span of less than about 1, less than about 0.8, less than about 0.7, less than about 0.65, or even less than about 0.5.
• the particles may have a size distribution with a span in a range of from about 0.1 to 1, from about 0.2 to 0.8, or from about 0.3 to 0.75.
• the surface-modifying agents of the present disclosure may include any molecules capable of coupling to particles useful for bioseparation (e.g., via covalent interactions, ionic interactions, hydrophobic interactions, or combinations thereof), and following such particle coupling, coupling to one or more analytes (e.g., via covalent interactions, ionic interactions, hydrophobic interactions, or combinations thereof) such that the analytes may be separated from a sample.
• the surface-modifying agents of the present disclosure may include at least a binding segment, a linking segment, and a reactive segment:
• the binding segment may include any segment capable of bonding the surface-modifying agent to the particles.
• the bond may be achieved, for example, covalently, hydrophobically, ionically, or combinations thereof.
• the binding segment may include a silyl group, e.g., binding segments having a formula:
• each group R 1 includes independently OH ⁇ or a hydrolyzable group from among halo, alkoxy, aryloxy, aralkyoxy, and acyloxy;
• each group R 2 includes independently a non-hydrolyzable group from among alkyl, aryl, and aralkyl.
• the linking group may include any segment suitable for connecting the binding segment with the reactive segment.
• the linking segment may comprise alkylene, arylene, or both, and optionally further comprises —NH— or alkyleneoxy, or both.
• the reactive segment may include any segment capable of coupling to one or more analytes such that the analyte may be separated from a sample (e.g., a solution having an analyte dispersed therein).
• the reactive segment may include a reactive nitrogen group, e.g., reactive segments having a formula:
• each group R 3 comprises independently hydrogen, alkyl, aryl, or aralkyl
• the surface-modifying agent may include (aminoethylaminomethyl)phenethyltrimethoxysilane (SIA0588.0, available from Gelest, Inc., Tullytown, Pa.), N-(2-aminoethyl)-3-aminopropylmethyldimethoxysilane (SIA0589.0, Gelest), N-(2-aminoethyl)-3-aminopropyltrimethoxysilane (SIA0591.0, Gelest), N-(6-aminohexyl)aminopropyltrimethoxysilane (SIA0594.0, Gelest), N-(2-aminoethyl)-11-aminoundecyl-trimethoxysilane (SIA0595.0, Gelest), aminophenyltrimethoxysilane (SIA0599.2, Gelest), N-3[(amino(polypropylenoxy)]aminopropyltrimethoxysilane (SIA059
• the present disclosure may relate to a method of making bioseparation compositions (e.g., the bioseparation compositions described above).
• the method may include size fractionating a first volume of particles to yield a second volume of particles having a desired particle size distribution. Any conventional size fractionation method may be employed including filtration, decantation, sedimentation, centrifugation, wet or dry screening, air or liquid elutriation, cyclones, static electricity, or combinations thereof.
• size fractionating yields a second volume of particles having a particle size distribution that is narrower than the particle size distribution of the first volume. Size fractionating may yield a second volume of particles having a particle size distribution with a span of less than about 1, less than about 0.8, less than about 0.7, less than about 0.6, or even less than about 0.5.
• the method may further include surface modifying at least a portion of the particles.
• surface modifying may include subjecting the particles to an optional pre-treatment step (e.g., to expose or clean a surface of the particle to facilitate surface modification).
• the optional pre-treatment step may include an alkaline treatment.
• the pretreatment may include an acid or plasma cleaning treatment.
• the method may include introducing the particles into a treatment solution that includes one or more surface-modifying agents and one or more solvents, thereby producing particles having the surface-modifying agents bonded to or otherwise exhibited on the exterior surface of the particles (i.e., surface-modified particles).
• the surface-modification treatment solution may be agitated (stirred, shaken, etc.) and/or temperature controlled (e.g., heated) to facilitate surface modification of the particles.
• the surface-modification treatment solution may include one or more solvents (e.g., organic and inorganic liquids (including water) or plasticizers known to be used or useful to dissolve or soften other organic or inorganic materials) and one or more surface-modifying agents.
• the treatment solution may include at least 0.01 wt. %, at least 0.04 wt. %, at least 0.1 wt. %, at least 1.0 wt. %, at least 4.0 wt. %, or even at least 10 wt. % of surface-modifying agent, based on the total weight of the surface-modification treatment solution.
• the ratio of surface-modification treatment solution to particles may vary widely. This ratio, and the concentration of the surface-modifying agent in the treatment solution, can be used to control the amount of modifying agent that is coupled to the particles. Generally, the amount of surface-modification treatment solution may be at least enough to wet the surface of the particles, or may be enough to allow for dispersion of the particles in the solution, thus allowing for agitation during the treatment process. In illustrative embodiments, the ratio of surface-modification treatment solution to particles may be at least 0.25:1, at least 0.5:1, at least 1:1, at least 2:1, at least 5:1, or even at least 10:1, on a volume basis.
• the surface-modified particles may be separated from the treatment solution (e.g., by filtration), optionally washed, and dried.
• the present disclosure may relate to a method for capturing an analyte in a solution.
• the method may include providing surface-modified particles, and contacting the surface-modified particles with a solution having one or more analytes capable of coupling to (or otherwise capable of being captured by) the surface-modifying agent, thereby generating surface-modified particles carrying an analyte.
• the method may further include agitating (e.g., inverting, stirring, shaking, etc.) the solution to achieve dispersion of the surface-modified particles throughout the solution.
• the solution may be agitated such that the surface-modified particles are substantially uniformly dispersed throughout the solution.
• the method may also include separating the analyte-carrying surface-modified particles from the solution.
• separating the particles from the solution may include allowing the analyte carrying surface-modified particles to float to an upper surface (i.e., air/solution interface) of the solution.
• the surface-modified hollow particles of the present disclosure may facilitate rapid separation of the particles from the solution.
• the particles of the present disclosure may separate in less than about 2 minutes, less than about 1 minute, less than about 30 seconds, or even less than about 15 seconds. In this manner, all manner of analytes may be readily captured by the surface-modified particles of the present disclosure.
• the method may include capture of one or more proteins and/or one or more nucleic acids from a solution utilizing the surface-modified particles of the present disclosure.
• compositions and methods of the present disclosure may be useful in, for example, food pathogen detection, medical diagnostics, cell separation, and environmental monitoring and/or cleaning.
• fractionation interval 5-10 g of glass bubbles and 400 mL of water were added to a cylindrical separatory funnel, mixed well by inverting the funnel up and down 3-4 times, and secured upright for a period (a “fractionation interval”) prior to removal of the liquid phase.
• the glass bubbles separated into three segments: (i) a top segment floating at the surface of the water in the funnel; (ii) a bottom segment of fines and shards that settled out of the water; and (iii) a middle segment forming a cloudy suspension of water and glass bubbles between the top and bottom segments.
• the fractionation interval was selected such that these three distinct segments were apparent and was determined empirically for the desired particle size distribution for a given bubble density and diameter.
• the fractionation interval was selected to be relatively short (e.g., 1 minute) in order to drain and remove the bubbles of segments (ii) and (iii). That is, the fractionation period was selected such that it was shorter than the time necessary for the glass bubbles of segment (ii) to rise to the surface of the water in the funnel.
• Particle size distribution of the glass bubbles was measured by light scattering using a laser particle analyzer (SATURN DIGISIZER 5200; Micromeritics; Norcross, Ga.).
• the fraction of the particles for which 10 percent of the particles in the distribution were smaller (10P) were determined from the particle size distribution.
• Example 5 a sample of GB1 was fractionated in accordance with the Fractionation Procedure.
• Separation time was determined by mixing 0.05 mL of each sample with 3 mL of DI water in a glass vial (14 mm inner diameter, water height of 30 mm), and placing upright to allow the suspension to separate. Separation time, determined as the time for substantially all of the glass bubbles to float to the top surface of the water, was measured. The results are shown in Table 3.
• GB1 glass bubbles were fractionated in accordance with the Fractionation Procedure. Approximately 3 grams of the fractionated glass bubbles were added to 100 mL of 0.01 N or 0.1 N sodium hydroxide in a glass jar. The jar was placed on a shaker for at least 6 hours and the bubbles were drained and dried. For Examples 6-11, 0.5 g of the alkaline treated glass bubbles were added to 20 g of a mixture of 95 wt % ethanol and 5 wt % water, and the pH was adjusted to 5.0 with acetic acid.
• Aminosilane modified, fractionated glass bubbles from Examples 6-11 and unmodified, fractionated glass bubbles from Example 1 were tested for DNA capture by mixing 10 milligrams of the glass bubbles with 1.5 mL of 9 ⁇ g/mL of calf thymus DNA (Sigma-Aldrich) in PBS (Phosphate buffered saline, pH 7.0, Sigma) in a centrifuge tube. The tubes were inverted up and down repeatedly for 1 minute at room temperature. The tubes were placed upright for about 10 seconds for the glass bubbles to float to the surface, leaving a clear solution underneath.
• the clear solution was collected and tested for absorbance measurement at 260 nm using a spectrophotometer (Agilent 8453 UV-VIS spectrophotometer; Agilent Technologies; Santa Clara Calif.) to determine the amount of DNA in the sample after adding glass bubbles as compared to a reference sample before adding the bubbles.
• the amount of captured DNA is shown in Table 5 in micrograms of DNA per milligram of glass bubbles.
• Example preparation (ug/mg-glass bubble) 20
• Example 6 0.330 21
• Example 7 0.413 22
• Example 8 0.473 23
• Example 9 0.158 24
• Example 10 0.360 25
• Example 11 0.435 R3
• Example R1 ⁇ 0.008 R4 Example R2 0.008
• Example 20 milligrams of an amine-coated fractionated glass bubbles from Example 12 were added to a mixture of 1.5mL of 1 mg/mL BSA (Bovine serum albumin; Sigma-Aldrich; St. Louis, Mo.) in 10 millimolar PBS buffer with the pH was adjusted to 6.0, 7.0 or 8.0, using 0.1 N HCl or 0.1 N NaOH and mixing in a centrifuge tube. Tubes were inverted up and down repeatedly for 2 minutes at room temperature. The tubes were placed upright for approximately 10 seconds for the glass bubbles to float to the surface and leave a clear solution underneath. The clear solution was collected for absorbance measurement at 280 nm using an Agilent 8453 UV-VIS spectrophotometer.
• Example R5 was prepared with unmodified, fractionated K25 glass bubbles from Example R1.

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