EP1183327A1 - Filtres a chelateurs de metaux et filtres a chelates de metaux - Google Patents

Filtres a chelateurs de metaux et filtres a chelates de metaux

Info

Publication number
EP1183327A1
EP1183327A1 EP00926556A EP00926556A EP1183327A1 EP 1183327 A1 EP1183327 A1 EP 1183327A1 EP 00926556 A EP00926556 A EP 00926556A EP 00926556 A EP00926556 A EP 00926556A EP 1183327 A1 EP1183327 A1 EP 1183327A1
Authority
EP
European Patent Office
Prior art keywords
filter
metal
linker
species
groups
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Withdrawn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
EP00926556A
Other languages
German (de)
English (en)
Inventor
Edward Hanna Kachab
Graeme Ross Barnett
Martin Smith
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.)
Abbott Rapid Diagnostics Pty Ltd
Whatman International Ltd
Original Assignee
Panbio Pty Ltd
Whatman International Ltd
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
Application filed by Panbio Pty Ltd, Whatman International Ltd filed Critical Panbio Pty Ltd
Publication of EP1183327A1 publication Critical patent/EP1183327A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
    • B01D67/0081—After-treatment of organic or inorganic membranes
    • B01D67/0093—Chemical modification
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
    • B01D69/14—Dynamic membranes
    • B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
    • 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
    • B01J45/00—Ion-exchange in which a complex or a chelate is formed; Use of material as complex or chelate forming ion-exchangers; Treatment of material for improving the complex or chelate forming ion-exchange properties

Definitions

  • the present invention relates generally to the separation and analysis of molecules.
  • the invention relates to the isolation of these molecules from a mixture in a fluid phase on a porous membrane comprising a metal chelating filter or metal chelate filter
  • porous membrane is in a microplate format, tube, vial
  • bioaffinity molecules may then be analysed using a wide variety of methodologies including the employment of specific bioaffinity molecules, specific
  • the molecules may be released
  • Microporous membranes as support matrices have also
  • Ligand immobilised membranes provide a compact
  • Ligands have been immobilised on support materials in the
  • membranes have been initially coated with a water-insoluble protein such as Zein,
  • Porous membrane materials used for non-covalent ligand immobilisation in affinity chromatography have included materials such
  • PVDF polyvinylidene fluoride
  • Non-covalent binding results in random orientation of the ligand and can
  • peptides and the oligonucleotides are not only covalently bound to the
  • peptides may be coupled to a solid support via
  • biomolecules as ligands onto activated membranes are:-
  • N ⁇ 2+ and Zn 2+ are the most widely.
  • Suitable donor ligands include ammo acids histidine, cysteine and tryptophan Since natural proteins rarely contain suitably arranged donor
  • ligands recombinant proteins are engineered with a metal chelate
  • binding tag at either the C- or N- terminal ends of the protein for
  • microporous filter materials such as Sepharose (IDA-sepharose, Pharmacia), magnetic beads (Ni-NTA Magnetic Agarose Beads, Qiagen) and microtitre plates (Ni-NTA HisSorb, Qiagen).
  • Iminodiacetic acid (IDA) has also been covalently attached to a stable IDA
  • MPS microporous plastic sheet
  • the MPS matrix is an inert polymeric microporous sheet that contains
  • silica that can either be fu ⁇ ctionalised with ion exchange groups or affinity ligands solely for the purpose of protein purification.
  • metal chelate filter or metal chelating species filter when processed in accordance with the invention to provide a metal chelate filter or metal chelating species filter may provide a number of
  • the invention provides a ligand immobilisation
  • the invention provides for the capture and
  • the invention has particularity in the diagnostics, high-throughput screening, and biotechnology industries.
  • the invention has particularity in the diagnostics, high-throughput screening, and biotechnology industries.
  • the invention has particularity in the diagnostics, high-throughput screening, and biotechnology industries.
  • the invention has particularity in the diagnostics, high-throughput screening, and biotechnology industries.
  • the invention has particularity in the diagnostics, high-throughput screening, and biotechnology industries.
  • the process of the invention includes the following variants:-
  • the invention includes within its scope products produced
  • filter means a porous material
  • filter media having an average pore size in the region of 0.01 to 1000 microns and more preferably 0.1-5 microns.
  • the filter may be formed from either fully or partly from
  • accessible functional groups such as -Si-OH groups or which may be treated with a reagent to provide accessible functional groups including -OH, -Si-OH, -NH 2 and other amine
  • groups including alkylamino, thiols, cyanobromides, aldehydes, carboxylates, sulfonylchlorides, ketones, halogens,
  • haloacetyl inclusive of chloroacetyl, iodoacetyl or bromoacetyl and
  • epoxy groups may be attached directly to the filter or by an appropriate spacer arm or linker.
  • porous filter materials comprising
  • polyamides inclusive of nylon, cellulose acetate, nitrocellulose,
  • polyvinylidene fluoride or other fluoropolymers, polysulfone, paper, or
  • linkers and/or a metal chelating species or a metal chelate may also be suitable media for the attachment of linkers and/or a metal chelating species or a metal chelate.
  • linker as used herein means any spacer group
  • the linker prior to use has an appropriate functional group as
  • linker can be utilised which may include substituted or
  • unsubstituted alkyl groups having from one to twenty, or more preferably one to six carbons, and wherein the alkyl groups may be linear or
  • branched Linkers may also comprise substituted or unsubstituted aryl or aryl alkyl groups
  • the linking group may be variable comprising a single methylene or a plurality of methylene groups
  • linking group may also comprise peptides or branched peptides inclusive
  • linkers include, for example, both linear,
  • polyamides polyethyleneimines, polyarylene sulfides, polysiloxanes,
  • polyimides polyacetates, dend ⁇ mers and dendrime ⁇ c-like molecules or
  • linker as used
  • cross-linking species or reagents used to couple the metal chelate or metal chelating species to accessible functional
  • cross-linking species either on the filter or which have been produced by de ⁇ vatisation
  • Such cross-linking species are referred to by way of example to the
  • microfibre In a preferred embodiment of the invention, microfibre
  • glass filters made from silica are especially adapted for use in the
  • Such filters have silanol (Si-OH) functional groups as part of
  • the filter matrix may be derivatised using a suitable reagent to
  • such reagents may have an in-built linker or the linker may be pre-attached to the metal
  • the metal chelating species may be any metal chelating species.
  • the metal chelating species includes a functional group capable of reacting with Si-OH groups such as triethoxysilane or trimethoxysilane.
  • silanisation reagents which may be used for the
  • filters comprise accessible amine groups on the surface of the filter.
  • linker or metal chelating species with linker attached may
  • peptides as linkers.
  • filters may be treated with a reagent such as ethylenediamine which may be
  • a short linker in a suitable solvent such as acetonitrile and usually in the presence of a catalyst such as triethylamine.
  • Suitable reactive groups may need to be
  • Such hydroxyl groups can be directly used for the attachment of a filter
  • hydroxyl groups may be further derivatised with silanisation reagents
  • Such reagents for example may include cyanogen bromide, tosyl chloride, N-hydroxy succinamide, diamines, hydrazine and its
  • dialdehydes such as glutaraldehyde, carbodiimides, and
  • MCS metal chelating species
  • metal chelating species include iminodiacetic acid, nitrilotriacetic acid, diethylenetriamine
  • metal chelate means the metal chelating species having a metal coordinated thereto. Chelation type associations make use of metal ions such as but not limited to, the
  • metal chelate filters of the invention which include the
  • metal can be used for immobilisation of ligands that coordinate with the
  • species may be attached to the filter by a covalent bond, charge
  • metal chelating species may be attached to the filter via a linker by either of the
  • the linker may be pre-attached to the metal chelating species followed by attachment of
  • suitable protecting groups include Fmoc, Boc, trityl groups and any other protecting groups.
  • branched species such as lysine or branched polymeric species such as dendrimers may increase
  • FIG. 1 shows a derivatised glass fibre filter
  • imidodiacetic acid metal chelate attached to the filter with a linker.
  • FIG. 2 shows a polyacrylamide gel with a low range
  • dendrimer-HRP conjugate at about 73 kDa is readily apparent in the
  • FIG. 3 shows results of an experiment using Zn 2+ immobilised on the filter in which HRP, HRP-antibody conjugate and HRP-dendrimer conjugate were passed through metal chelate filter discs
  • FIG. 4 shows free HRP, HRP-antibody and E5 dendrimer-
  • metal chelate glass fibre filters (Whatman GF/B, GF/C and GF/F).
  • metal ion used was Zn 2+ .
  • FIG. 5 shows results of an experiment in which E5-
  • dendrimer conjugate, diluent only, E5 dend ⁇ mer only and E5 dendrimer- antibody conjugate were passed through Zn 2+ metal chelate filter discs
  • FIG 6 of a polyacrylamide gel shows that metal chelate filter discs efficiently capture a 6 x histidine tagged protein from a
  • the concentration of the tagged protein was found to be approximately five times higher in material eluted from the discs than material that flowed through the discs
  • TNBS trmitrobenzynesulfonic acid
  • HBTU Benzotriazole-N,N,N',N',-Tetramethyl-Uronium-Hexafluorophosphate
  • HOBt N-Hydroxybenzothazole
  • DIPEA diisopropylethylamine
  • the Fmoc group was then tested with a TNBS test.
  • the deprotected filters were treated with nitrilotriacetic acid
  • Nitrilotriacetic acid derivatised filter discs were each placed on a sintered glass funnel connected to a vacuum pump. The discs were
  • FIG. 1 shows an example of a molecular structure showing the derivatised filter, linker(s) and the metal chelate.
  • FIG. 2 shows examples of different molar ratios of E5 dendrimer-HRP conjugations.
  • FIG. 3 shows results of an experiment using Zn 2+
  • HRP-dendrimer conjugate were passed through metal chelate filter discs
  • FIG. 4 shows free HRP, HRP-antibody and E5 dendrimer-
  • metal chelate glass fibre filters (Whatman GF/B, GF/C and GF/F).
  • E5-dendrimer conjugate, diluent only, E5 dendrimer only and E5 dendrimer-antibody conjugate were passed through Zn 2+ metal
  • TMB substrate was applied and washed as described above.
  • FIG. 5 shows the results. It can be seen that the presence
  • FIG 6 of a polyacrylamide gel shows that metal chelate filter discs efficiently captured a hexahistidme tagged protein from a
  • MC and MCS filters may be incorporated into an immunoassay filter plate, column, syringe filter housing or tube for a wide
  • the MC and MCS filters may also be embodied with a second type filter such as an FTATM Gene Guard
  • the MC and MCS filters may further embodied with a second filter such as to restrict
  • the bound genetic material such as human genomic DNA can be ultimately removed by use of reagents such as restriction endonucleases
  • the fragments of the restriction digest can be incubated in the presence of a MC filter harbouring
  • probes could be derivatised at their 5' or 3' ends with a metal chelate tail such as a hexahistidme tag or dendrimer e g PAMAM
  • genomic DNA E g traditional methodology (other than FTA) of purification yields genomic DNA as a soluble fraction After restriction digest, or in vitro
  • template nucleic acid can be produced that is in a position to be captured by such
  • oligonucleotide probes may be mixed with the genomic DNA whereby the
  • DNA/probe hybrids formed can be captured on MC
  • a second probe conjugated to a detector label e g
  • Oligonucleotides, mRNA and cDNA coupled to dendrimers or other species capable of acting as a ligand, for example a hexahistidine tag, can be
  • dendrimers or a hexahistidine may be added directly to a MC filter sheet to form a microarray suitable for analysis without the need for a multi-well plate format.
  • proteins including a hexahistidine tail may be immobilised on a metal chelate filter. This will
  • Unstable recombinant protein may be immobilized on MC filters quickly and at room temperature
  • incorporating an MC filter would enable the user to rapidly immobilize potential drugs or target
  • the method could also be combined with a FTA filter for simultaneous
  • MCF-captured proteins may then be tested for function in bioassays.
  • An indicator of cellular events is the change in kinase activity. Interaction at the cell surface is
  • a synthetic kinase substrate is incubated with the kinase sample of
  • the substrate was engineered to contain a 6xHis
  • a MC filter will have greater specificity than Promega's streptavidin filter that is commonly used together with biotin tagged peptides.
  • the isolated library compound may then be further interrogated to
  • target molecule may be conjugated to a dendrimer or hexahistidine tag either of which
  • dendrimer or hexahistidine may then be further
  • Overlapping peptide sequences may be tagged with a hexahistidme tail or dendrimers and immobilised onto metal chelate filters for subsequent screening
  • metal chelate filter bottoms could be used to capture hexahistidine or dendrimer tagged or target molecules
  • product may be engineered to bear a hexahistidme sequence This product may then readily be captured onto a metal chelate filter
  • a deep well multiwell plate encapsulating a MC filter at the bottom of each well may be constructed
  • each well may be carried out mammalian, yeast or bacterial culture without fluid dripping through the filter bottoms Protein
  • FTA and MC filter materials within the same device may be constructed.
  • the cells will lyse, the cells
  • episomal and/or genomic DNA will bind to the FTA, and the recombinant protein containing an
  • the recombinant protein can be harvested and assayed. If the protein collected is of interest
  • Metalloproteases are implicated with the onset of
  • metastasis being part of the mechanism utilised by
  • Recombinant proteins and enzymes expressing a tag such as a hexahistidine tail may be assayed directly from culture to determine a) levels of
  • each tube, column or well will amplify the capacity
  • microorganisms or parts of microorganisms such as antigens or nucleic acids

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Inorganic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Manufacturing & Machinery (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)

Abstract

L'invention concerne un procédé de fabrication d'un filtre à chélate de métaux ou un filtre à espèces chélatrices de métaux, qui consiste en ce qui suit: traiter un filtre possédant une taille de pores entre 0,01 et 1000 micromètres et les groupes fonctionnels accessibles avec un chélate de métaux ou des espèces chélatrices de métaux pour obtenir un filtre à chélate de métaux ou un filtre à espèces chélatrices de métaux.
EP00926556A 1999-05-14 2000-05-15 Filtres a chelateurs de metaux et filtres a chelates de metaux Withdrawn EP1183327A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
AUPQ0343A AUPQ034399A0 (en) 1999-05-14 1999-05-14 Metal chelating filters and metal chelate filters
AUPQ034399 1999-05-14
PCT/AU2000/000477 WO2000070012A1 (fr) 1999-05-14 2000-05-15 Filtres a chelateurs de metaux et filtres a chelates de metaux

Publications (1)

Publication Number Publication Date
EP1183327A1 true EP1183327A1 (fr) 2002-03-06

Family

ID=3814547

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00926556A Withdrawn EP1183327A1 (fr) 1999-05-14 2000-05-15 Filtres a chelateurs de metaux et filtres a chelates de metaux

Country Status (3)

Country Link
EP (1) EP1183327A1 (fr)
AU (1) AUPQ034399A0 (fr)
WO (1) WO2000070012A1 (fr)

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US9610546B2 (en) 2014-03-12 2017-04-04 Lockheed Martin Corporation Separation membranes formed from perforated graphene and methods for use thereof
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US9744617B2 (en) 2014-01-31 2017-08-29 Lockheed Martin Corporation Methods for perforating multi-layer graphene through ion bombardment
US10653824B2 (en) 2012-05-25 2020-05-19 Lockheed Martin Corporation Two-dimensional materials and uses thereof
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CN105940479A (zh) 2014-01-31 2016-09-14 洛克希德马丁公司 使用宽离子场穿孔二维材料
CN106029596A (zh) 2014-01-31 2016-10-12 洛克希德马丁公司 采用多孔非牺牲性支撑层的二维材料形成复合结构的方法
CA2942496A1 (fr) 2014-03-12 2015-09-17 Lockheed Martin Corporation Membranes de separation formees a partir de graphene perfore
CA2973472A1 (fr) 2014-09-02 2016-03-10 Lockheed Martin Corporation Membranes d'hemodialyse et d'hemofiltration basees sur un materiau membranaire bidimensionnel et procedes employant ces dernieres
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JP2018530499A (ja) 2015-08-06 2018-10-18 ロッキード・マーチン・コーポレーション グラフェンのナノ粒子変性及び穿孔
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KR102357806B1 (ko) 2021-08-10 2022-02-09 주식회사 에이치이공일 친환경 금속 킬레이트 겔필터, 이의 제조방법 및 이를 이용한 연수 샤워 헤드
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CN115583710A (zh) * 2022-07-19 2023-01-10 江苏驰佳环保科技有限公司 一种螯合剂及其制备方法

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Also Published As

Publication number Publication date
WO2000070012A1 (fr) 2000-11-23
AUPQ034399A0 (en) 1999-06-03

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