EP1525299A2 - Support de reseau de petales pour microplaques - Google Patents

Support de reseau de petales pour microplaques

Info

Publication number
EP1525299A2
EP1525299A2 EP03771827A EP03771827A EP1525299A2 EP 1525299 A2 EP1525299 A2 EP 1525299A2 EP 03771827 A EP03771827 A EP 03771827A EP 03771827 A EP03771827 A EP 03771827A EP 1525299 A2 EP1525299 A2 EP 1525299A2
Authority
EP
European Patent Office
Prior art keywords
petal
ion
shaped
shaped purification
purification members
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP03771827A
Other languages
German (de)
English (en)
Inventor
Paul Ramstad
Michael Harrold
Kevin Hennessy
Aldrich Lau
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Applied Biosystems Inc
Original Assignee
Applera Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US10/414,179 external-priority patent/US20040018559A1/en
Priority claimed from US10/413,935 external-priority patent/US6833238B2/en
Application filed by Applera Corp filed Critical Applera Corp
Publication of EP1525299A2 publication Critical patent/EP1525299A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02—Making microcapsules or microballoons
    • B01J13/06—Making microcapsules or microballoons by phase separation
    • B01J13/14—Polymerisation; cross-linking
    • 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
    • B01J47/00—Ion-exchange processes in general; Apparatus therefor
    • B01J47/018—Granulation; Incorporation of ion-exchangers in a matrix; Mixing with inert materials
    • 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
    • B01D15/34—Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
    • 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
    • G01N1/4005—Concentrating samples by transferring a selected component through a membrane
    • G01N2001/4016—Concentrating samples by transferring a selected component through a membrane being a selective membrane, e.g. dialysis or osmosis

Definitions

  • an apparatus having a petal-array of purification materials that can be disposed within respective wells of a multi-well microplate, for example, a standard-format 96- or 384- well plate. Methods of making and using the apparatus are also provided.
  • the purification material can be located on an array of members, for example, petal-shaped purification members, adapted for insertion into a corresponding array of reaction wells.
  • the purification members can also include binding sites for target components.
  • An apparatus and method for facilitating the release of labeled monomers from a purification and binding support within a microplate format are also provided.
  • an analyte-manipulation apparatus is provided.
  • the apparatus can include, for example, a plurality of wells defining an array, wherein each of the wells includes a rim defining an opening at an upper end of each well, with the openings being disposed within a first plane.
  • the apparatus can include a support, for example, a sheet, including a p lurality o f petal-shaped p urification o r i on-exchange m embers formed t herein a t positions corresponding to the wells of the array, with the support being disposed along a second plane above and substantially parallel to the first plane, and with at least one of the petal-shaped purification members being positioned near each one of the openings.
  • the apparatus can include a stack of supports, for example, formed as individual sheets, disposed above the well openings, with each support of the stack including a plurality of petal-shaped purification members integrally formed therein, and with each petal-shaped purification member of each support being disposed at a position corresponding to a respective one of the wells of the array.
  • the stack of supports can include more than one support, for example, at least three of the supports, for example, 3, 4, 5, 6, 7, 8, 9, 10, or more supports.
  • Each of the petal-shaped purification members can be movable between (i) a first position, substantially within the second plane, and (ii) a second position, at least partially disposed outside of the second plane and extending at least partially into a nearby well via a respective opening.
  • the apparatus can further include a platen including a major surface facing the support, and a plurality of ring-shaped projections extending outwardly from the major surface of the platen.
  • the platen can be adapted for movement toward and away from the support, whereby upon moving the platen toward the support, the projections can pressingly engage the petal-shaped purification members, thereby deflecting the petal-shaped purification members from the first position to the second position.
  • each of the ring-shaped projections can taper in a direction away from the major surface.
  • the platen and each of the ring-shaped projections of the platen defines a passage extending longitudinally through each ring-shaped projection and through the platen.
  • An instrument for example, a pipette, can be inserted through the passage to access the interior region of any one or more of the wells when the petal- shaped purification members are deflected into their respective wells.
  • a sample and/or reagent can be deposited into or withdrawn from one or more selected wells by using an instrument via the passage.
  • the apparatus can further include a die plate disposed between the support and the plurality of wells, wherein the die plate includes an array of apertures extending therethrough, with each of the apertures being disposed at a position corresponding to a respective one of the wells of the array.
  • Figures 1A and IB are partial side-sectional views of an apparatus according to various embodiments
  • Figure 2 is an exploded, perspective view of the apparatus shown in Figure 1A; [00011] Figure 3 i s a t op p Ian v iew s howing a s upport i ncluding p etal-shaped p urification members, according to various embodiments;
  • Figures 4A and 4B are enlarged top plan views showing a plurality of petal-shaped purification members, each taken from a respective support of an aligned stack of eight supports, individually and superposed, respectively;
  • Figures 5 A and 5B show a die plate, according to various embodiments, in top plan view and side elevational view, respectively;
  • Figures 6 A and 6 B s how a p laten, a ccording t o v arious e mbodiments, i n t op p Ian view and side elevational view, respectively;
  • Figure 7B is a cross-sectional view through different lines, of the SEJE particle shown in Figure 7A and used as a purification material according to various embodiments.
  • size-exclusion ion-exchange (SEIE) particles having an ion-exchange core micro-encapsulated by a shell capable of size-exclusion are provided.
  • SEIE size-exclusion ion-exchange
  • a core of liquid, solid, and/or g as i s m icro-encapsulated w ith a s hell to e ontrol a ccess t o t he c ore.
  • micro-encapsulation can coat the entire exterior surface of the core (and optionally interior surfaces), or it can coat only a portion of the exterior surface of the core (and optionally interior surfaces).
  • micro-encapsulation of the core can be irreversible to permanently coat the core, or reversible to release the core upon dissolution of the coating.
  • micro-encapsulation can include encapsulation of an agglomerate of core material in a shell.
  • the aggregate can be fused, sintered, pressed, compressed, or otherwise formed together core materials.
  • the core material can be a single particle and not an aggregate.
  • core or “core material” can refer to a single particle or an aggregate of particles.
  • shell refers to coating any portion of the core exterior surface and/or interior surface. The dimensions and formation of the shell are described below.
  • the term "material” refers to any substance on a molecular level or in bulk.
  • a material can be a liquid and/or solid, e.g. an emulsion or a resin.
  • an apparatus 10 can include one or more supports, for example, the stack of support sheets 12a-h.
  • the number of supports can be any suitable number, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.
  • each of the support sheets 12a-h independently can include a polymeric film, for example, a polycarbonate or polystyrene film, having a thickness of between about O.OOlinch and about 0.010 inch, for example, about 0.004 inch.
  • the film can be textured to increase its effective surface area.
  • each support sheet can be a die-cut, chemically-treated, membrane or film support.
  • Figure 3 shows a single support sheet, 12a, from the stack of support sheets 12a-h of Figure 2, in top plan view.
  • Support sheet 12a can be configured with outer dimensions generally like that of a top surface of a microplate with which the support sheet 12a is to be used.
  • Support sheet 12a can be die-cut to provide an array of members, for example, petal-shaped purification members 21a.
  • the members can be any suitable shape, for example, petal-shaped, rectangular, or finger-shaped.
  • the petal-shaped purification members 21a can be arranged in an array corresponding to an array of wells in the microplate 18 with which the support is to be used, for example, a regular rectangular array.
  • the petal-shaped purification members are arranged in a 12 X 8 array, with adjacently disposed petal-shaped purification members being spaced 0.9 cm center-to-center.
  • Other array configurations are contemplated herein, for example, a 24 X 16 array, with adjacently disposed petal-shaped purification members being spaced 0.45 cm center-to-center.
  • Other suitable configurations will be apparent to those of ordinary skill in the art upon review of the disclosure and/or practice of the present teachings as described herein.
  • Each of the support sheets 12a-h can include one or more location features to facilitate alignment with respect to other system components.
  • slots 22 can be formed at selected locations along the edge regions of each of the support sheets 12a-h.
  • the slots 22 can be positioned and configured to mate with complementary-shaped regions of one or more of the microplate 18, die plate 14, and platen 16.
  • Figure 2 shows a protrusion 26 capable of mating with the slots 22, wherein the protrusion 26 is formed at a mid-point along each edge region of the die plate 14.
  • all the petal-shaped purification members of any one of the supports can face, or "point," in the same direction.
  • the directionality of the petal- shaped purification members can differ between any two of the supports. That is, the petal- shaped purification members of any one support can point in a direction that differs from that of any of the other supports.
  • each support includes a petal-shaped purification member disposed at a position that is radially distinct from the petal-shaped purification members of the other supports of the stack.
  • Figure 4A shows petal-shaped purification members 21a-h from a selected coordinate, for example, row 1, column 1, of each of the eight supports 12a-h of the stack from Figure 2.
  • applying a downwardly-directed force against a petal-shaped purification member can deflect the member from its normal position to a second position, for example, below the plane of the support. Upon r emoving the force, the resilient petal-shaped purification member can return substantially to its normal position.
  • Figures 5 A and 5B show the die plate 14 in top plan and side elevational views, respectively.
  • the die plate 14 can include protrusions 26 for properly locating and aligning one or more supports thereon, for example, by way of slots 22 in supports 12a-h.
  • Use of location features c an facilitate 1 ocation o f an array o f p etal-shaped p urification members o f a s upport directly over respective well openings in a microplate.
  • the die plate 14 can include an array of holes or apertures 30 that are concentric, and directly correspond to, the wells of the microplate 18.
  • the die plate can include features that align it relative to the microplate 18, and/or to the platen 16.
  • FIGS 6A and 6B show the platen 16 in top plan and side elevational views, respectively.
  • the platen 16 can include passages or through-holes 34 that are concentric with and directly correspond to the wells of microplate 18. Except for such through-holes, the platen can be configured to substantially cover one or more support.
  • the platen 16 can include ring-shaped projections 36 extending from a major surface 16a, with each ring- shaped projection circumscribing, and further defining, a respective one of the through-holes 34.
  • Such construction can permit access to the individual wells of the microplate through the platen from a region extending above each of the wells of the microplate.
  • an outer circumferential region of each ring-shaped projection 36 of the platen 16 can be configured with a taper along a direction extending away from the major surface 16a of the platen 16 and extending towards supports 12a-h.
  • the taper can facilitate placement and seating of each ring-shaped projection 36 in a corresponding aperture 30 of the die plate 14 upon bringing the platen 16 and die plate 14 together, as shown in Figure IB and as described further below.
  • the platen 16 can include slots 38 as shown in Figure 6 A.
  • the slots 38 can have a shape similar to the slots 22 of the supports 12a-h. Slots 38 can assist in properly locating and aligning the platen 16 over the die plate 14 by mating the slots 38 of the platen 16 with the projections 26 of the die plate 14.
  • microplate 18 can be an injection molded plastic plate, the length and width of which conform to a commonly used standard, for example, a rectangle of 5.03 inches x 3.37 inches (127.8 mm by 85.5 mm).
  • wells are formed integrally with the microplate, and can be arranged, for example, in a 12 X 8 regular rectangular array and spaced 0.9 cm center-to-center.
  • the illustrated embodiments show arrangements configured in accordance with the popular 96-well format, the present teachings also contemplate any other number of wells, for example, 12, 24, 48, 192, or 384 wells, laid out in any suitable configuration, for example, square, rectangular, circular, ovoid, or other regular or irregular patterns.
  • Each of the petal-shaped purification members 21 can be moved between (i) a first position, outside of a corresponding well, and (ii) a second position, extending at least partially into the corresponding well.
  • a platen 16 can be placed over the stack of support sheets 12.
  • the platen 16 can include a major surface 16a facing the support sheets 12, and a plurality of ring-shaped projections 36 can extend outwardly from the major surface 16a toward the support sheets 12.
  • the platen 16 can be moved toward and away from the support sheets 12.
  • the projections 36 can pressingly engage the petal-shaped purification members 21, thereby deflecting the petal-shaped purification members 21 from the first to the second position, as depicted in Figure IB.
  • the ring-shaped projections 36 of the platen 16 can pressingly engage and deflect the petal-shaped purification members 21 of the support sheets 12 against the holes in the die plate 14 and into the wells of the microplate 18, where the petal- shaped purification members 21 can chemically interact with the contents of the individual wells.
  • one or more chemicals, biochemicals, or purification medium can be present on at least a portion of one or more of the petal-shaped purification members.
  • the petal-shaped purification members can be introduced into respective wells that can contain a first sample, such as a polymerized chain reaction product or DNA sequencing product.
  • the chemicals, biochemicals, and/or purification medium on the petal-shaped purification members can interact with the first sample to bind one or more components of the sample.
  • the chemicals, biochemicals, and/or purification medium on the petal-shaped purification members can bind desirable components, for example, DNA fragments, dsDNA, ssDNA, polynucleotides, oligonucleotides, and the like.
  • the chemicals, biochemicals, and/or purification medium on the petal-shaped purification members can bind undesirable reaction products, including fragments, salts, promoters, terminators, reactive dyes, and other undesirable reaction products as known to those of ordinary skill in the art.
  • the petal-shaped purification members can be chemically treated.
  • One or more of the petal-shaped purification members can include one or more biochemicals immobilized thereon.
  • biochemicals can include, for example, one or more nucleic acids.
  • biochemicals can include one or more DNA-sequencing reagents, such as terminators, primers, or a combination thereof.
  • At least a portion of the petal-shaped purification members can have a purification medium, for example, size-exclusion ion-exchange particles, ion-exchange particles, a size-exclusion resin, or a combination thereof, affixed thereto.
  • Figures 7 A and 7B are not drawn to scale, and the relation between objects in the figures, such as the relation between core pore sizes and shell pore sizes, is not to scale, and can in fact be inverse, such that the core pore size is larger than the shell pore size.
  • large molecules such as long single stranded DNA (ssDNA) fragments, and double stranded DNA (dsDNA)
  • ssDNA long single stranded DNA
  • dsDNA double stranded DNA
  • large molecules can slide past or bounce off an exterior surface 121 of the shell 120, and remain in solution rather than ion-exchanging with the anion-exchange particle 130.
  • Small molecules such as deoxynucleotide triphosphates (dNTPs), dye-labeled deoxynucleotide triphosphates, dideoxynucelotide triphosphates (ddNTPs), dye- labeled dideoxynucelotide triphosphates, and small ions, such as chlorine, can pass through the pores 125 of the size-exclusion shell 120 and can undergo ion-exchange with anion-exchange resin 113 at or near the interface 123 of the shell and ion-exchange core 111, or within the pores of ion-exchange core 111.
  • dNTPs deoxynucleotide triphosphates
  • ddNTPs dideoxynucelotide triphosphates
  • small ions such as chlorine
  • Figure 7A shows a partial cut-away 133 showing a surface of the ion-exchange core 111 coated with anion-exchange resin 113.
  • the anion-exchange resin 113 for example, a cross- linked, macroporous copolymer of methyl methacrylate and 2-hydroxy-3- methacryloyloxypropyltrimethylammonium chloride, can be present on all internal and external surfaces of the solid core material 112. Together, the anion-exchange resin and solid core material or support 112 form the anion-exchange core 111.
  • Counter-ions released from the anion-exchange core 111 can react with a counter-ion, for example, a counter-ion such as a hydroxide ion, of a cation-exchanger 135 that can be provided in a mixture with the SEIE particle 130, to produce a neutral molecule such as water.
  • a counter-ion for example, a counter-ion such as a hydroxide ion
  • a cation-exchanger 135 can be provided in a mixture with the SEIE particle 130, to produce a neutral molecule such as water.
  • SEIE particles can be used in a mixture, a mixed bed, or a homogeneous bed of particles. Wherein a homogeneous bed of anionic- or cationic-SEIE particles is used, the counter-ion can be released directly into a sample solution upon ion-exchange. In certain cases, the presence of the counter-ion in the sample solution does not affect further processing or reaction of the sample.
  • a purification device can include size-exclusion ion-exchange (SELE) particles embedded in a substrate.
  • the SEJE particles can be ion-exchange particles, as described elsewhere herein, encapsulated by a size-exclusion resin, as described elsewhere herein.
  • An ion-exchange particle can be encapsulated by a size-exclusion resin, for example, by inverse emulsification and polymerization, to form an SEIE particle.
  • an anionic ion-exchange particle can be formed by impregnating polyethyleneimine onto the surface of a solid core material, including all surfaces of the pores of the solid core material.
  • the resultant ion-exchange particle can be encapsulated by a shell of size-exclusion resin, for example, polyacrylamide, to form a size-exclusion anion-exchange (SEAE) particle.
  • SEAE size-exclusion anion-exchange
  • other anion-exchange resins or cation-exchange resins can be impregnated or retained on at least a portion of the internal surfaces, on at least a portion of the external surface, or on at least a portion of all surfaces of the solid core material of the ion-exchange particle.
  • a solid core material capable of ion-exchange can be encapsulated by a size-exclusion resin to form an SEJJE particle.
  • the ion-exchange particle can be surface- activated to enhance or aid in formation of the shell around the ion-exchange particle.
  • Surface activation of the ion-exchange particle can include, for example, derivatization of functional groups on the ion-exchange particle by monomers; absorption of polyanions onto the ion- exchange particle by ionic interaction with an ion-exchange resin of the ion-exchange particle; passive adsorption onto the ion-exchange particle of a neutral, water-soluble polymer; or adsorption of a charged initiator on the ion-exchange particle through ionic interaction.
  • Greater details about SELE particles are described in concurrently filed U.S. Patent Application No. 10/414,179 to Lau et al, entitled "Size-Exclusion Ion-Exchange Particles," Attorney Docket No. 4885US, which is incorporated herein in its entirety by reference.
  • a purification device in the form of a coated stick can be made, wherein the coated stick has SEIE particles embedded in a substrate.
  • the SEIE particles can be encapsulated, attached, adhered, embedded, or otherwise in contact with a substrate, for example, a polymeric substrate, by heat application, by heated extrusion of the substrate in the presence of the particles, by pressure, by physical force, by chemical treatment, by electrical current, by ultrasonication, by molding with the substrate, or by other methods of attachment known to those of ordinary skill in the art, or combinations thereof.
  • each support can be a single-layer film or membrane material.
  • Each support can include one or more registration feature, for example, one or more slot formed in each support, to facilitate alignment of the supports with respect to a microplate.
  • the petal-shaped purification members of each support can be resiliently deformable, tending to return to the first position after having been deflected therefrom.
  • a method for biochemical interaction can include immobilizing one or more selected biochemicals on a plurality of petal-shaped purification members, wherein the petal-shaped purification members are disposed in an array on a support.
  • One or more reagents can be introduced into a plurality of wells, wherein the wells are disposed in an array corresponding to the array of petal-shaped purification members.
  • the petal-shaped purification members can be positioned above the plurality of wells so that each petal-shaped purification member is situated above a corresponding one of the plurality of wells.
  • the petal- shaped purification members can be pressingly engaged from a side opposite the wells to simultaneously move the petal-shaped purification members into the corresponding wells, contacting the one or more selected biochemicals on the plurality of petal-shaped purification members with the one or more reagents in the plurality of wells, and permitting a biochemical interaction to take place involving the one or more selected biochemicals and the one or more reagents.
  • a method for biochemical interaction can include providing a plurality of petal-shaped purification members, wherein the petal-shaped purification members are disposed in an array on a support.
  • a microplate including a plurality of wells disposed in an array corresponding to the array of petal-shaped purification members can be provided, wherein one or more selected biochemicals can be placed in the wells of the microplate.
  • the petal-shaped purification members can be positioned above the microplate so that each petal-shaped purification member is situated above a corresponding one of the plurality of wells.
  • the petal-shaped purification members can be pressingly engaged from a side opposite the wells to simultaneously move the petal-shaped purification members into the corresponding wells to contact the one or more selected biochemicals therein.
  • One or more of the selected biochemicals in the wells can be immobilized on the petal-shaped purification members.
  • the petal-shaped purification members with the one or more selected biochemicals immobilized thereon can be withdrawn from the wells.
  • the method can further include providing a second microplate including a plurality of wells disposed in an array corresponding to the array of petal-shaped purification members, wherein one or more selected reagents is disposed in the wells of the second microplate.
  • the petal-shaped purification members with one or more biochemical immobilized thereon can be positioned above the second microplate so that each petal-shaped purification member is situated above a corresponding one of the plurality of wells.
  • the petal-shaped purification members can be pressingly engaged from a side opposite the second microplate to simultaneously move the petal-shaped purification members into the corresponding wells to contact the one or more reagents t herein.
  • a b iochemical i nteraction c an t ake p lace i n t he w ells i nvolving t he o ne o r more selected biochemicals immobilized on the petal-shaped purification members and the one or m ore s elected r eagents i n t he w ells.
  • a ccording t o v arious e mbodiments, t he p etal-shaped purification members can release the one or more immobilized selected biochemicals from the petal-shaped purification members into the wells.
  • a method for purifying a sample in one or more wells in a plurality of wells disposed in an array is provided.
  • a plurality of petal-shaped purification members disposed in an array on a support can be provided, wherein each of the petal-shaped purification members is at least partially coated with a purification medium, for example, ion-exchange particles, a size-exclusion resin, or size-exclusion ion-exchange particles.
  • the support can be positioned above the microplate such that each petal-shaped purification member is situated above a corresponding one of the plurality of wells.
  • the petal-shaped purification members can be pressingly engaged from a side opposite the wells to simultaneously move the petal-shaped purification members into the corresponding wells to contact the samples in the wells.
  • the petals can remain in contact with the respective samples for a period of time sufficient to remove at least 70%, at least 80%, or at least 90% of the impurities from the sample in the well.
  • the petal-shaped purification members with the impurities immobilized thereon can be withdrawn from the wells.
  • the purification material can adsorb or retain the desired portions of the sample, and leave the impurities in the well.
  • the support sheets containing the petal-shaped purification members with at least a portion of the first sample immobilized thereon can be moved individually or collectively into a position over a second microplate, wherein the petal-shaped purification members can be pressingly engaged by a platen such that the petal-shaped purification members are deflected into the respective wells of the second microplate, wherein the petal-shaped purification members can react with a second sample in the wells of the second microplate to release the immobilized components of the first sample.
  • the petal-shaped purification members can be used for purification of a sample. At least a portion of the petal-shaped purification members can be coated with a purification medium, for example, ion-exchange particles, a size-exclusion resin, or size-exclusion ion-exchange particles, to facilitate purification.
  • a purification medium for example, ion-exchange particles, a size-exclusion resin, or size-exclusion ion-exchange particles, to facilitate purification.
  • a purification medium for example, ion-exchange particles, a size-exclusion resin, or size-exclusion ion-exchange particles, to facilitate purification.
  • a purification medium for example, ion-exchange particles, a size-exclusion resin, or size-exclusion ion-exchange particles, to facilitate purification.
  • "resin” includes compositions with the characteristics of a resin or a gel.
  • an ion-exchange particle immobilized on a petal- shaped purification member can be an anionic or cationic material.
  • the ion-exchange particle can be a polymer, cross-linked polymer, or inorganic material, for example, silica.
  • the ion-exchange particle can be a solid core material capable of ion-exchange, or a solid core material treated with an ion-exchange resin.
  • the ion-exchange particle can be surface-activated.
  • the ion-exchange particle can be non-magnetic, paramagnetic, or magnetic.
  • Exemplary ion-exchange particle materials include Macro-Prep® ion-exchange resins from Bio-Rad, andNucleosil® silica-based ion-exchange resins from Macherey-Nagel.
  • the solid core material can be macroporous silica, controlled pore glass (CPG), a macroporous polymer microsphere with internal pores, other porous materials as known to one of ordinary skill in the art, or a combination thereof.
  • the solid core material can have various surface features, including, for example, pores, crevices, cracks, or depressions.
  • the solid c ore m aterial c an i n clude s odium o xide, s ilicon d ioxide, s odium b orate, o r a c ombination thereof.
  • the solid core material can be modified to be capable of ion-exchange, for example, cation-exchange or anion-exchange. Modification of the solid core material can include treatment of the solid core material to form cationic or anionic substituent groups on the surfaces of the solid core material.
  • the term "surface" can include an external surface and internal surfaces, for example, the surfaces of voids or pores within the solid core material.
  • the solid core material can be modified to include quaternarized functional groups, at least one carboxylic acid group, at least one sulfonic acid group, other cationic or anionic functional groups known to one of ordinary skill in the art, or a combination thereof on the surface of the solid core material.
  • the solid core material can be porous, microporous, or macroporous.
  • the solid core material can have an average pore size of less than or equal to 1000 Angstroms, from 100 Angstroms to 1000 Angstroms, or less than or equal to 100 Angstroms.
  • the average diameter of the solid core material can be from 0.1 ⁇ m to 100 ⁇ m, from 1 ⁇ m to 50 ⁇ m, or from 2 ⁇ m to 20 ⁇ m, according to various embodiments.
  • the average diameter of the solid core material can be 100 ⁇ m or less, 50 ⁇ m or less, or 20 ⁇ m or less.
  • a solid core material can adsorb an ion-exchange resin onto the external surface, internal surface, or both the external and the internal surface of the solid core material to form an ion-exchange particle.
  • the ion-exchange resin can be a cation- exchange resin or an anion-exchange resin.
  • the ion-exchange resin can include quatemarized functional groups, at least one carboxylic acid group, at least one sulfonic acid group, or a combination thereof. Suitable anion-exchange resins and cation-exchange resins are known to one of ordinary skill in the art.
  • the ion-exchange capacity of the ion- exchange particle can be improved by increasing a mass of ion-exchange resin, such as quaternary ammonium resin, on the external surface and/or on the internal surfaces of the pores of the solid core material of the ion-exchange particle.
  • the ion-exchange capacity of the ion- exchange particle can be improved by selection of cationic or anionic functional groups on the external surface, internal surfaces of the pores, or both internal surfaces and external surface of the solid core material.
  • the ion-exchange resin can be fo ⁇ ned in situ on the solid core material.
  • the ion-exchange resin can be the product of one or more monomer, one or more polymer, or a combination thereof, according to various embodiments.
  • a solid core material of SiO 2 having an average pore size of 1000 Angstroms, a void volume of 0.95cc/g, and a diameter of 5 ⁇ m, can be added to a solution of polyethyleneimine in methanol and incubated for a time sufficient to impregnate the polyethyleneimine on all internal and external surfaces of the solid core material.
  • polyethyleneimine can be adsorbed due to hydrogen bonding with silanol groups in the solid core material.
  • the adsorbed polyethyleneimine can be reacted with a second monomer, such as, for example, 1,3-dibromopropane, in a solvent, for example, dioxane, followed by placement in water, to form a quaternary ammonium gel that functions as an anion-exchange resin on the solid core material, forming an ion-exchange particle.
  • a second monomer such as, for example, 1,3-dibromopropane
  • solvent for example, dioxane
  • Other methods can be used and are described, for example, in concurrently filed U.S.
  • Such methods include methods used to impregnate or retain other anion-exchange resins or cation-exchange resins on at least a portion of the internal surfaces, on at least a portion of the external surface, or on at least a portion of all surfaces of the solid core material of the ion-exchange particle.
  • purification of a sample with an ion-exchange particle can be achieved by ion-exchange. Displacement of counter-ions from the ion-exchange / particle during ion-exchange can release a large number of counter-ions into a sample solution.
  • anionic ion-exchange particles and cationic ion-exchange particles can both be present during purification of a sample such that counterions of the ion- exchange particles react to form a neutral molecule, for example, water.
  • the ion-exchange particles can both be present on a petal-shaped purification member, or at least one of cationic or anionic ion-exchange particles can be present, for example, in the sample.
  • the ion-exchange particles can be placed in a monomer solution including at least one p olymerizable p olymer and o ne o r m ore o f a c atalyst, i nitiator, c ross-linker, c hain s topper, surfactant, terminator, promoter, buffer, accelerator, or a combination thereof.
  • the monomer solution can be capable of polymerizing or cross-linking at about room temperature.
  • the monomer solution can be capable of polymerizing upon application of heat, application of radiation, addition of a catalyst, addition of an initiator, or a combination thereof.
  • a petal-shaped purification member can be inserted into the monomer solution including the ion-exchange particles, and the monomer solution can be polymerized and/or cross-linked to adhere the ion-exchange particles to the petal- shaped purification member.
  • Other methods of attaching the ion-exchange particles to the petal- shaped purification members will be apparent to those of ordinary skill in the art.
  • the monomer solution can be capable of forming a size-exclusion resin.
  • a size-exclusion resin with or without ion-exchange particles therein can be coated on at least a portion of one or more of the petal-shaped purification members.
  • a size-exclusion resin can be a cross-linked product of two or more reactive monomeric units.
  • the monomeric units can be water-soluble monomeric units.
  • water-soluble includes materials with any degree of water solubility from slightly water-soluble to highly water-soluble, and materials that are swellable in water.
  • the monomeric units can be nifrogen-containing, oxygen-containing, or both.
  • the size- exclusion resin can be a homopolymer or a copolymer.
  • the size-exclusion resin can be the reaction product of an acrylamide, such as polyacrylamide, polymethylmethacrylate, polyethylmethacrylate, or polymethacrylate.
  • the size-exclusion resin can be a reaction product of acrylamide and N,N'- methylenebisacrylamide.
  • exemplary water-soluble polymers can include, but are not limited to, polyacrylamide, poly(N-methylacrylamide), poly(N,N- dimethylacrylamide), poly(N-vinylformamide), poly(N-vinylacetamide), poly(N-methyl-N- vinylacetamide), poly(vinylalcohol), poly(hydroxyethyl (meth)acrylate), poly(vinypyrrolidone), methylcellulose, ethylcellulose, other suitable polymers capable of cross-linking as known to one of ordinary skill in the art, or a combination thereof.
  • the size-exclusion resin can be neutral, anionic, or cationic. According to various embodiments, the size-exclusion resin can be hydrophilic.
  • the size-exclusion resin can be a cross-linked polymer network of polymers capable of swelling in water, for example, hydrogels. Exemplary hydrogels are described, for example, in U.S. Patent No. 6,380,456 Bl, inco ⁇ orated herein in its entirety by reference.
  • the size-exclusion resin, once formed, can be non-water soluble. According to various embodiments, the size- exclusion resin can prevent adso ⁇ tion of ssDNA fragments and/or double-stranded DNA (dsDNA) fragments.
  • the size-exclusion resin can be formed with pores of a pre-determined size.
  • the pores can be of the same or varying size.
  • the pores can function as the size-exclusion factor for preventing molecules larger than a certain size from passing through the size-exclusion resin to the ion-exchange core which is encapsulated by the size-exclusion resin.
  • the size-exclusion resin can be formed by cross-linking one or more reactive monomer by addition of a cross-linker, for example, N,N'-methylenebisacrylamide, a free-radical initiator, a chain-stopper, a surfactant, a catalyst, a terminator, a promoter, a buffer, an accelerator, or a combination thereof.
  • a cross-linker or initiator can be added in an amount of from 1.0 mol% to 95 mol%.
  • the cross-linker or initiator can be added in an amount of from 1.0 mol% to 50 mol%, from 2.0 mol% to 20 mol%, from 5 mol% to 15 mol%, or from 10 mol% to 12 mol%.
  • the amount of cross-linker or initiator used to form the size- exclusion resin is at least one factor in determining the size of the pores of the size-exclusion resin, and the size-exclusion ability of the size-exclusion resin.
  • the choice of cross-linker or initiator, and/or selection of the reaction conditions can control the amount of cross-linking of the size-exclusion resin.
  • various multifunctional cross- linkers can be used that have varying amounts of functionaUty.
  • the appropriate amount of a cross- linker to use to form a desired size-exclusion resin pore size can be determined by those of ordinary skill in the art based on the functionaUty of the cross-linker chosen, the reaction conditions, and other factors as known to those of ordinary skill in the art.
  • the monomer solution can be capable of polymerizing or cross-linking at about room temperature.
  • the monomer solution can be capable of polymerizing upon application of heat, application of radiation, addition of a catalyst, addition of an initiator, or a combination thereof.
  • the resultant size-exclusion resin can have a pore size equal to or smaller than a 10 nt ssDNA, or equal to or smaller than a 100 nt ssDNA.
  • the r esultant i on- exchange particle can be encapsulated by a shell of size-exclusion resin, for example, polyacrylamide, to form a size-exclusion anion-exchange (SEAE) particle.
  • size-exclusion resin for example, polyacrylamide
  • SEIE size-exclusion anion-exchange
  • other anion-exchange resins or cation-exchange resins can be impregnated or retained on at least a portion of the internal surfaces, on at least a portion of the external surface, or on at least a portion of all surfaces of the solid core material of the ion-exchange particle.
  • a solid core material capable of ion-exchange can be encapsulated by a size-exclusion resin to form an SEIE particle.
  • the ion-exchange particle can be surface- activated to enhance or aid in formation of the shell around the ion-exchange particle.
  • Surface activation of the ion-exchange particle can include, for example, derivatization of functional groups on the ion-exchange particle by monomers; abso ⁇ tion of polyanions onto the ion- exchange particle by ionic interaction with an ion-exchange resin of the ion-exchange particle; passive adso ⁇ tion onto the ion-exchange particle of a neutral, water-soluble polymer; or adso ⁇ tion of a charged initiator on the ion-exchange particle through ionic interaction.
  • a size-exclusion resin shell can be formed on an ion-exchange particle wherein the shell has a thickness of from 2% to 300% of the diameter of the ion-exchange particle.
  • the shell can have a thickness of from 25% to 200% of the diameter of the ion-exchange particle.
  • the thickness of the shell can vary over the surface of the ion-exchange particle, or the thickness of the shell can be uniform over the surface of the ion-exchange particle.
  • the shell can at least partially encapsulate the ion-exchange particle.
  • the shell material can at least partially fill one or more pore or surface feature, for example cracks, of the ion-exchange particle.
  • all internal and external surfaces of the ion-exchange particle can be coated with a monomer suitable for forming a shell.
  • the monomer can be reacted with one or more of a second monomer, a catalyst, or an initiator to form a cross-linked size-exclusion polymer on all surfaces of the ion-exchange particle, with the outermost surface of the size-exclusion polymer forming a shell.
  • Large molecules can not contact the ion-exchange particle because of the presence of the polymer forming the size-exclusion shell on all surfaces of the ion-exchange particle.
  • the ion-exchange particle can be a solid core material coated with an ion-exchange resin.
  • counter-ions released from the ion-exchange particle can react with the counter-ion of an ion-exchanger to produce a neutral molecule, such as water.
  • the ion- exchanger can be present on the petal-shaped purification member, or in the sample solution.
  • the ion-exchanger can be another SEIE particle.
  • the size-exclusion resin can be used for purification o f biological samples, for example, PCR products, to separate larger DNA, for example dsDNA, from ssDNA, free nucleotides, and salts.
  • the pores of the size- exclusion resin or shell of a SEIE particle have a pore size capable of excluding a molecule equal to or larger than 10 nt ssDNA, salts and nucleotides, for example, present in a sample to be purified, can pass through the size-exclusion resin.
  • 50% or more of the pores of the size-exclusion resin h ave a p ore s ize c apable o f e xcluding a m olecule e qual t o o r 1 arger t han 10 n t, t he s ize- exclusion resin can be used for purification of biological samples, for example, from a sequencing reaction. Purification of a sequencing reaction sample can remove dye-labeled dideoxynucleotides and salts from the sequencing reaction sample, leaving a purified sample containing ssDNA in an amount of 70% or more, 80% or more, 90% or more, or 95% or more of the eluted sample volume.
  • a sample for purification c an b e a P CR p roduct solution containing, for example, buffer salts, metal ions, polymerase, nucleotides, oligonucleotide primers, and other components.
  • PCR products can be used in subsequent enzymatic reactions that can be sensitive to at least some of the artifacts found in a sample solution containing the PCR products.
  • free nucleotides and oligonucleotide primers can interfere with downstream enzymatic reactions.
  • a size-exclusion resin or SEIE particle in a purification device can have a pore size capable of excluding a molecule equal to or larger than a 100 nt ssDNA, allowing nucleotides, oligonucleotide primers less than about 100 nt in size, and buffer salts to pass through the size-exclusion resin. 100 nt or larger molecules can remain in the sample solution.
  • At least 50% or more of the surface of the size-exclusion resin or SEIE particle shell is capable of excluding a molecule equal to or larger than a 100 nt ssDNA, and allowing nucleotides, oligonucleotide primers, and buffer salts less than 100 nt in size to pass through the size-exclusion resin, or though the shell of the SEIE particle to the ion-exchange core.
  • the resulting purified sample solution can contain purified PCR products in a desalted environment, and can be used in downstream reactions and analyses.
  • PCR purification can be directed toward purifying larger dsDNA separate from smaller ssDNA, free nucleotides, and salts.
  • PCR product purification using a petal-shaped purification member having one or more of ion- exchange particles, a size-exclusion resin, or SEIE particles affixed thereto can isolate a 250-600 bp amplicon, can remove 44 nt primers and/or nucleotides, or both isolate and remove.
  • a sample for purification can be a DNA sequencing reaction solution containing, for example, buffer salts, metal ions, enzymes,polymerase, nucleotides, oligonucleotide primers, and other components. Purification of sequencing reaction solutions can have different requirements than purification of PCR reaction solutions.
  • finished sequencing reactions can contain residual dye-labeled dideoxynucleotides (terminators) that can be removed prior to electrophoretic analysis and DNA sequencing or basecalling. Failure to remove terminators can result in "blobs" that can cause enors in DNA sequencing or basecalling.
  • capillary sequencers can use electrokinetic injection as a means to introduce DNA sequencing reaction samples. The presence of salts in the samples can effect the introduction of the sample into the capillary.
  • DNA sequencing reaction samples can be highly desalted by purification with ion-exchange particles, a size-exclusion resin, or SEIE particles.
  • a sample solution purified as described herein can have a salt connection less than or equal to about 100 ⁇ M, or less than or equal to 50 ⁇ M.
  • a purified sample solution can be suitable for electrokinetic capillary injection. T he s ize-exclusion r esin o r shell o f the S EIE p article c an b e c apable o f e xcluding a molecule equal to or larger than a 10 nt ssDNA, allowing smaller molecules, for example, salts and nucleotides, in a sample solution to pass through the size-exclusion resin or shell.
  • about 50% or more of the surface of the size-exclusion resin or SEIE shell can be capable of excluding a molecule equal to or larger than about a 10 nt ssDNA, allowing smaller molecules, for example, salts and dye-labeled nucleotides, to be removed from the sample solution, and leaving ssDNA free in the sample solution.
  • a sequencing reaction purification can separate ssDNA, for example, ssDNA of from about 10 nt to about 800 nt in size, from, for example, dye-labeled nucleotides and salts.
  • the upper portions of the microplate wells can be configured to act in place of the die plate, thus eliminating the die plate from the above- described assembly.
  • Many varieties of microplates, available from different suppliers, can be accommodated by the inco ⁇ oration of a spring-loaded centering means within the basic assembly.
  • the present teachings provide a method of delivering to or removing from a sample well salts, dyes, DNA, terminators, primers, or other components useful for, or resulting from, PCR or DNA sequencing; a facility for the exposure of one or more discreet supports to an anay of individual samples; and, a tool for moving and/or transferring large numbers of labeled samples at a time.
  • the present teachings further provides the ability to capture nucleotides or other biological samples on multiple binding sites on a support within a standard laboratory microplate format, and enables subsequent release of the nucleotides or other biological samples from the support, also within the microplate format.
  • the present teachings further provides a method of purifying a PCR product or DNA sequencing reaction product in bulk.
  • the present teachings provide a plurality of discreet, labeled, petal-shaped purification members and their support in a single deformable film or membrane. This design can eliminate many handling and alignment issues associated with stacking and placing many post/array assemblies with densely packed probes, for example, twelve, into the distinct wells, for example, 96, of a standard microtiter plate.
  • All publications and patent applications refened to herein are hereby inco ⁇ orated by reference in their entirety, and to the same extent as if each individual publication or patent application was specifically and individually indicated to be inco ⁇ orated by reference.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Manufacturing Of Micro-Capsules (AREA)
  • Devices For Use In Laboratory Experiments (AREA)

Abstract

L'invention porte sur des dispositifs comportant des supports pouvant recevoir un ou des matériaux échangeurs d'ions en vue de la purification d'échantillons. Dans les différentes exécutions les supports comprennent des éléments déformables, par exemple des éléments de purification en forme de pétales, constituant des sites de fixation du matériau échangeur d'ions, et facultativement de substances biologiques, de produits chimiques, de sels, ou autres. L'invention porte également sur un appareil et des procédés d'introduction et de retrait d'éléments de purification dans les différents puits d'une microplaque à puits multiples.
EP03771827A 2002-07-26 2003-07-25 Support de reseau de petales pour microplaques Withdrawn EP1525299A2 (fr)

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
US39885202P 2002-07-26 2002-07-26
US398852P 2002-07-26
US10/414,179 US20040018559A1 (en) 2002-07-26 2003-04-14 Size-exclusion ion-exchange particles
US10/413,935 US6833238B2 (en) 2002-01-04 2003-04-14 Petal-array support for use with microplates
US414179 2003-04-14
US413935 2003-04-14
US10/413,797 US20040016702A1 (en) 2002-07-26 2003-04-14 Device and method for purification of nucleic acids
US413797 2003-04-14
PCT/US2003/023253 WO2004011592A2 (fr) 2002-07-26 2003-07-25 Support de reseau de petales pour microplaques

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EP03771854A Withdrawn EP1525053A1 (fr) 2002-07-26 2003-07-25 Dispositif et procede de purification d'acides nucleiques
EP03771827A Withdrawn EP1525299A2 (fr) 2002-07-26 2003-07-25 Support de reseau de petales pour microplaques

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Families Citing this family (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6627159B1 (en) 2000-06-28 2003-09-30 3M Innovative Properties Company Centrifugal filling of sample processing devices
US7192560B2 (en) 2001-12-20 2007-03-20 3M Innovative Properties Company Methods and devices for removal of organic molecules from biological mixtures using anion exchange
US7347976B2 (en) 2001-12-20 2008-03-25 3M Innovative Properties Company Methods and devices for removal of organic molecules from biological mixtures using a hydrophilic solid support in a hydrophobic matrix
US7981600B2 (en) 2003-04-17 2011-07-19 3M Innovative Properties Company Methods and devices for removal of organic molecules from biological mixtures using an anion exchange material that includes a polyoxyalkylene
US7322254B2 (en) 2003-12-12 2008-01-29 3M Innovative Properties Company Variable valve apparatus and methods
US7727710B2 (en) 2003-12-24 2010-06-01 3M Innovative Properties Company Materials, methods, and kits for reducing nonspecific binding of molecules to a surface
US7939249B2 (en) 2003-12-24 2011-05-10 3M Innovative Properties Company Methods for nucleic acid isolation and kits using a microfluidic device and concentration step
EP2277987A3 (fr) * 2004-02-18 2011-10-12 Life Technologies Corporation Particules échangeuses d'ions revétues de polyelectrolytes
SE0400490D0 (sv) * 2004-02-26 2004-02-26 Amersham Biosciences Ab Plasmid purification
JP2008536478A (ja) * 2005-02-15 2008-09-11 アプレラ コーポレイション 高分子電解質被覆イオン交換粒子
EP1848793B1 (fr) * 2005-02-15 2012-06-20 Life Technologies Corporation Particules d'echange ionique revetues de polyelectrolytes
US7378260B2 (en) * 2005-04-01 2008-05-27 Applera Corporation Products and methods for reducing dye artifacts
US7763210B2 (en) 2005-07-05 2010-07-27 3M Innovative Properties Company Compliant microfluidic sample processing disks
US7323660B2 (en) 2005-07-05 2008-01-29 3M Innovative Properties Company Modular sample processing apparatus kits and modules
US7754474B2 (en) 2005-07-05 2010-07-13 3M Innovative Properties Company Sample processing device compression systems and methods
US7935518B2 (en) * 2006-09-27 2011-05-03 Alessandra Luchini Smart hydrogel particles for biomarker harvesting
DE102007005655A1 (de) 2007-01-31 2008-08-07 Qiagen Gmbh Vorrichtung und Verfahren zur Aufreinigung von Nukleinsäuren
JP2012500651A (ja) * 2008-08-25 2012-01-12 ジーイー・ヘルスケア・バイオサイエンス・コーポレイション ゲノムｄｎａの精製のための簡単な負荷および溶離プロセス
US8834792B2 (en) 2009-11-13 2014-09-16 3M Innovative Properties Company Systems for processing sample processing devices
CN102958602A (zh) 2010-07-09 2013-03-06 通用电气健康护理生物科学股份公司 层析方法及为此使用的介质
EP2638156B1 (fr) 2010-11-09 2016-01-27 Qiagen GmbH Procédé et dispositif pour l'isolement et la purification d'acides nucléiques double brin
AU2012255151B2 (en) 2011-05-18 2015-09-03 Diasorin Italia S.P.A. Systems and methods for detecting the presence of a selected volume of material in a sample processing device
WO2012158988A1 (fr) 2011-05-18 2012-11-22 3M Innovative Properties Company Systèmes et procédés de distribution sur un dispositif de traitement d'échantillons
US8931331B2 (en) 2011-05-18 2015-01-13 3M Innovative Properties Company Systems and methods for volumetric metering on a sample processing device
US20120291869A1 (en) * 2011-05-20 2012-11-22 Dober Chemical Corporation Systems and Methods for Releasing Additive Compositions
EP2537415A1 (fr) * 2011-06-24 2012-12-26 David Vachon Compositions efficaces sur le plan biologique, articles manufacturés et procédés de production et/ou d'utilisation associés
US8822554B2 (en) * 2011-10-04 2014-09-02 Purolite Corporation Aminated ion exchange resins and production methods thereof
SG11201407808XA (en) 2012-05-31 2014-12-30 Agency Science Tech & Res Chromatographic Purification Of Polynucleotides With Negatively Charged Particles
GB201214193D0 (en) * 2012-08-08 2012-09-19 Ge Healthcare Uk Ltd Polymerase chain reaction method for amplifying nucleic acid
CN108384780B (zh) * 2018-02-11 2023-12-29 中国农业科学院烟草研究所 一种批量快速提取dna的方法及提取装置
JP7194129B2 (ja) * 2020-01-16 2022-12-21 株式会社豊田中央研究所 冷却液
NL2025044B1 (en) 2020-03-04 2021-10-14 Stichting Wetsus European Centre Of Excellence For Sustainable Water Tech Electrochemical device, system and method for electrochemically recovery and/or regeneration of carbon dioxide from a stream
JP7480070B2 (ja) 2021-01-15 2024-05-09 株式会社豊田中央研究所 冷却液

Family Cites Families (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4086217A (en) * 1976-03-03 1978-04-25 Hoffmann-La Roche Inc. Carcinoembryonic antigens
JPS57140644A (en) * 1980-12-15 1982-08-31 Asahi Chem Ind Co Ltd Molded product of composite adsorbent
AU642444B2 (en) * 1989-11-30 1993-10-21 Mochida Pharmaceutical Co., Ltd. Reaction vessel
CA2063855C (fr) * 1991-04-03 1997-08-26 Will Bloch Precision et exactitude de la separation d'acides nucleiques par echange d'anions
SE9700769D0 (sv) * 1997-03-04 1997-03-04 Pharmacia Biotech Ab Matriser för separation och separation som utnyttjar matriserna
EP1177043A2 (fr) * 1999-04-29 2002-02-06 Genome Therapeutics Corporation Dispositif permettant de traiter rapidement un echantillon d'adn par manipulation de liquide, thermocyclage et purification
EP1214149A2 (fr) * 1999-09-21 2002-06-19 Genome Therapeutics Corp. Dispositif de traitement rapide d'echantillons d'adn a manipulation des liquides, thermocyclage, et purification integres
AU1622501A (en) * 1999-11-23 2001-06-04 Porex Corporation Immobilized ion exchange materials and processes for making the same
SE9904272D0 (sv) * 1999-11-25 1999-11-25 Amersham Pharm Biotech Ab A method for selective removal of a substance from samples containing compounds having nucleic acid structure
US6284117B1 (en) * 1999-12-22 2001-09-04 Nanogen, Inc. Apparatus and method for removing small molecules and ions from low volume biological samples
US6504021B2 (en) * 2000-07-05 2003-01-07 Edge Biosystems, Inc. Ion exchange method for DNA purification
EP1379641B1 (fr) * 2000-12-20 2005-06-29 ALTANA Pharma AG Procede de production et de purification de proteines et de production de reseaux de proteines
US20020168643A1 (en) * 2001-05-08 2002-11-14 Wierzbowski Jamey M. Devices and methods for simultaneous separation and purification of molecular samples
KR100425536B1 (ko) * 2001-07-16 2004-03-30 학교법인 포항공과대학교 유체 마이크로칩용 브레드보드
US20030098271A1 (en) * 2001-11-26 2003-05-29 Ralph Somack Capsule and tray systems for combined sample collection, archiving, purification, and PCR
US7192560B2 (en) * 2001-12-20 2007-03-20 3M Innovative Properties Company Methods and devices for removal of organic molecules from biological mixtures using anion exchange

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2004011592A2 *

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AU2003254174A1 (en) 2004-02-16
WO2004011592A3 (fr) 2004-04-22
EP1525052A1 (fr) 2005-04-27
AU2003254175A1 (en) 2004-02-16
WO2004011592A2 (fr) 2004-02-05
AU2003256787A8 (en) 2004-02-16
WO2004011142A1 (fr) 2004-02-05
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EP1525053A1 (fr) 2005-04-27
AU2003256787A1 (en) 2004-02-16

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