WO2003004536A2 - Procede de fabrication de polysaccharides reticules, et leur utilisation en tant que phases stationnaires - Google Patents

Procede de fabrication de polysaccharides reticules, et leur utilisation en tant que phases stationnaires Download PDF

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Publication number
WO2003004536A2
WO2003004536A2 PCT/EP2002/007236 EP0207236W WO03004536A2 WO 2003004536 A2 WO2003004536 A2 WO 2003004536A2 EP 0207236 W EP0207236 W EP 0207236W WO 03004536 A2 WO03004536 A2 WO 03004536A2
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WO
WIPO (PCT)
Prior art keywords
nanoparticles
crosslinkers
stationary phases
separation processes
cross
Prior art date
Application number
PCT/EP2002/007236
Other languages
German (de)
English (en)
Other versions
WO2003004536A3 (fr
Inventor
Joachim Teller
Jutta Nagel
Michael Frank
Fritz Westphal
Original Assignee
Micromod Partikeltechnologie Gmbh
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.)
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Publication date
Application filed by Micromod Partikeltechnologie Gmbh filed Critical Micromod Partikeltechnologie Gmbh
Publication of WO2003004536A2 publication Critical patent/WO2003004536A2/fr
Publication of WO2003004536A3 publication Critical patent/WO2003004536A3/fr

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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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/262—Synthetic macromolecular compounds obtained otherwise than by reactions only involving carbon to carbon unsaturated bonds, e.g. obtained by polycondensation
    • 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
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/265—Synthetic macromolecular compounds modified or post-treated polymers
    • B01J20/267—Cross-linked polymers
    • 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
    • B01J20/28009—Magnetic properties
    • 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
    • B01J20/282—Porous sorbents
    • B01J20/285—Porous sorbents based on polymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0036—Galactans; Derivatives thereof
    • C08B37/0039—Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose
    • 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
    • B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
    • B01J2220/54—Sorbents specially adapted for analytical or investigative chromatography

Definitions

  • the invention relates to a process for the preparation of crosslinked polysaccharides which can be used in the form of spherical particles as stationary phases in separation processes.
  • Cross-linked polysaccharides in the form of spherical particles already have a number of established technical, biotechnological and biomedical applications.
  • particles whose matrix consists of cellulose, agarose or dextran are considered bioinert and are therefore unmodified, activated or functionalized for various chromatographic processes for the separation of predominantly biological-chemical structures.
  • Affinity chromatography has the largest share.
  • Such methods are used to separate or purify peptides and proteins, antibodies, immunoglobulins and enzymes, biochemically relevant polysaccharides and lipids, steroids, nucleic acids and related structures as well as differently structured toxins.
  • Biomedical applications of such particles are in the fields of dialysis or apheresis or are practiced in methods for extracorporeal detoxication. Furthermore, gel filtration, gel chromatography and ion exchange chromatography as well as related processes of desalination or decontamination of heavy metals or radionuclides are important technical fields of application for cross-linked polysaccharide particles.
  • crosslinkers have also been used for special applications: polyfunctional acylation reagents (PERRIER et al: US Pat. No. 6,132,750), hydrophilic polyalkylene glycol derivatives (WEISSLEDER et al .: US Pat. No. 5,514,379), organopolysiloxanes (YAMAZAKI et al .: US 5,658,849) and titanium or zirconium compounds (COTTRELL et al .: US 5,532,350).
  • polyfunctional acylation reagents PRORIER et al: US Pat. No. 6,132,750
  • hydrophilic polyalkylene glycol derivatives WEISSLEDER et al .: US Pat. No. 5,514,379
  • organopolysiloxanes YAMAZAKI et al .: US 5,658,849
  • titanium or zirconium compounds COTTRELL et al .: US 5,532,350
  • the porosity of polysaccharide particles is essentially checked via the type of crosslinking or mediated by the chemical structure of the crosslinking agent used.
  • LARSSON (US 5 723 601) claims a bead structure with diffusion and flow pores and HJERTEN et al. (US 5 135 050) a particle design with reduced porosity.
  • the present invention was therefore based on the object of making crosslinked polysaccharides accessible which are already structurally stabilized in the primary crosslinking.
  • the activating and functionalizing reagents as well as other functionalities e.g. fluorescence, magnetizability
  • nanoparticles which have reactive chemical sequences on the surface are used as a structuring primary crosslinker and a further molecular crosslinker for crosslinking the polysaccharides.
  • the type of crosslinking according to the invention can in principle be used for all types of polysaccharides, in particular that relates to dextran, chitosan, alginic acid, cellulose, starch and agarose, which can optionally be substituted or otherwise modified.
  • this networking is of particular interest in connection with the production of spherical particles with diameters between 1 ⁇ m and 1 mm and the use of agarose as the matrix material.
  • the nanoparticles with chemically reactive surfaces used for crosslinking are not limited by the diameter, the particle matrix or their surface chemistry. It has proven to be advantageous to use nanoparticles with diameters between 1 and 800 nm. Both inorganic macromolecular structures, such as polysilicic acid or titanium dioxide, and natural or synthetic organic macromolecular substances, for example polyvinyl compounds, polyesters or amides or silicones, can be used in the matrix materials thereof. It is also possible to use composite materials for the matrix of the crosslinking nanoparticles. Appropriate colored, fluorescent or magnetizable nanoparticles can be used as particulate crosslinkers for special fields of application, which also require visualization or the magnetic mobility of the crosslinked polysaccharide materials.
  • Suitable particle types which may have to be subjected to a further chemical surface modification, are described by TELLER et al. (EP 1 036 763) or by GRÜTTNER et al. (J. Magn. & Magn. Mater. 194, 8 (1999)).
  • the nanoparticles which act as particulate crosslinkers, have reactive sequences on the surface which can add or from the hydroxyl groups of the polysaccharide these are substituted.
  • reactive sequences on the surface which can add or from the hydroxyl groups of the polysaccharide these are substituted.
  • These are, for example, heterocumule sequences, such as isocyanates or isothiocyanates, activated alkene structures, for example with vinylsulfonyl residues or ring sequences from the field of small carbocycles or heterocycles, such as epoxides or aziridines.
  • Substitutable sequences have haloalkyl groups, acyl halides or hetero-analogous structures derived therefrom, such as 4,6-dichloro-1,3,5-triazinyl units.
  • the molecular crosslinkers used according to the invention are bifunctional or polyfunctional compounds which have chain lengths of 2 to 12 atoms between the binding sites.
  • the bi- or polyfunctional compounds contain reactive sequences as have already been described for the particulate crosslinkers. It is not necessary, but has proven to be advantageous if the molecular crosslinkers have the same functional groups as the reactive nanoparticles.
  • the structural analogy between the matrix of the particulate crosslinkers and sequences of the molecular crosslinkers can also have a favorable effect.
  • crosslinked polysaccharides according to the invention can be further functionalized chemically by methods or synthetic methods known per se.
  • the present method thus offers the possibility of producing crosslinked polysaccharides which contain both a particulate and a molecular crosslinker.
  • Polysaccharides crosslinked in this way can be used in the form of particles (beads) with diameters in the range from 1 ⁇ m to 1 mm as stationary phases in separation processes. Such separation processes are used in chemistry and biochemistry, biotechnology and other life science technologies, as well as biomedicine.
  • the particles produced according to the invention can be used in all customary media, solvents and buffers.
  • the beads can be sterilized using conventional methods.
  • these stationary phases can carry different reactive or affine groups on their surface or such structures are generated or converted during their use.
  • stationary phases can be equipped with visualizable or mobile properties according to the separation task by using colored, fluorescent or magnetizable nanoparticles as crosslinkers in their manufacture.
  • visual properties it is also possible to bind corresponding dyes or fluorescent dyes to the matrix of the stationary phases by known processes via addition or substitution reactions.
  • the reaction of the agarose with the crosslinking agent particles is carried out by stirring the reaction mixture for eight hours at a temperature of 50 ° C. and increasing the temperature to 80 ° C. for a further hour.
  • 9.41 oil peanut oil, Henry Laotte GmbH thermostatted to 85 ° C. is introduced and the agarose suspension is metered into the preheated oil with stirring at 700 rpm using a thin jet.
  • the emulsion is then cooled to ⁇ 40 ° C. and a suspension of agarose particles is formed with a maximum size distribution of around 100 ⁇ m.
  • the crude product is washed with petroleum ether (101), with ethanol (91), with a 50% (v / v) ethanol / water mixture (51) and finally with deionized water (17, 51).
  • DCT 2,4-dichloro-1, 3,5-triazinyl groups
  • A 10g Glycidether 100, Carl Roth GmbH) * Abs .: 569nm, Em.:585 nm) **** Sigma Aldrich Chemie GmbH
  • B 10g 1,3-bis (3-glycidyloxypropyl) tetramethyldisiloxane) *) ** Abs .: 485nm, em .: 510nm
  • C 4g trichloroisocyanuric acid) ****

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Materials Engineering (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Processes Of Treating Macromolecular Substances (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Compositions Of Macromolecular Compounds (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Abstract

L'invention concerne un procédé d'obtention de particules réticulées pouvant être utilisées comme phases stationnaires dans des procédés de séparation. La présente stratégie de réticulation est basée sur une réticulation primaire structurée, dotée de nanoparticules réactives et sur l'utilisation d'un autre réticulant moléculaire. En ce qui concerne les domaines d'application visant, entre autre, une visualisation ou la mobilité magnétique des matériaux polysaccharides réticulés, on peut utiliser, comme réticulants particulaires, des nanoparticules correspondantes colorées, fluorescentes ou magnétisables. Cette réticulation est toutefois d'un intérêt particulier en ce qui concerne la production de particules sphériques de diamètre compris entre 1 νm et 1 mm, et l'utilisation d'agarose comme matériau pour la matrice.
PCT/EP2002/007236 2001-07-03 2002-07-01 Procede de fabrication de polysaccharides reticules, et leur utilisation en tant que phases stationnaires WO2003004536A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10132131A DE10132131A1 (de) 2001-07-03 2001-07-03 Verfahren zur Herstellung vernetzter Polysaccharide und deren Verwendung als stationäre Phasen in Trennverfahren
DE10132131.7 2001-07-03

Publications (2)

Publication Number Publication Date
WO2003004536A2 true WO2003004536A2 (fr) 2003-01-16
WO2003004536A3 WO2003004536A3 (fr) 2003-03-20

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DE (1) DE10132131A1 (fr)
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007050017A1 (fr) * 2005-10-28 2007-05-03 Ge Healthcare Bio-Sciences Ab Support de séparation ayant différentes fonctionnalités
CN102335595A (zh) * 2010-07-20 2012-02-01 华东理工大学 一种新型的具有梯度分布的整体柱固定相的制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2714671A1 (fr) * 1994-01-05 1995-07-07 Conservatoire Nal Arts Metiers Nouveaux dérivés polysaccharidiques et leur emploi pour la préparation de phases stationnaires chirales utiles pour la séparation des isomères de composés chimiques.
US5587467A (en) * 1993-06-22 1996-12-24 Daicel Chemical Industries, Ltd. Separating agent for optical isomers and process for producing the same
EP1036763A1 (fr) * 1999-02-12 2000-09-20 micromod Partikeltechnologie GmbH Procédé de préparation de particules d'acide polysilicique colorées et fluorescentes

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5587467A (en) * 1993-06-22 1996-12-24 Daicel Chemical Industries, Ltd. Separating agent for optical isomers and process for producing the same
FR2714671A1 (fr) * 1994-01-05 1995-07-07 Conservatoire Nal Arts Metiers Nouveaux dérivés polysaccharidiques et leur emploi pour la préparation de phases stationnaires chirales utiles pour la séparation des isomères de composés chimiques.
EP1036763A1 (fr) * 1999-02-12 2000-09-20 micromod Partikeltechnologie GmbH Procédé de préparation de particules d'acide polysilicique colorées et fluorescentes

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ARSHADY R: "MICROSPHERES AND MICROCAPSULES: A SURVEY OF MANUFACTURING TECHNIQUES. PART 1: SUSPENSION CROSS-LINKING" POLYMER ENGINEERING AND SCIENCE, BROOKFIELD CENTER, US, Bd. 29, Nr. 24, Dezember 1989 (1989-12), Seiten 1746-1758, XP000826592 *
CORDULA GR]TTNER ET AL.: "New types of silica-fortified magnetic nanoparticles as tools for molecular biology applications" JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS, Bd. 194, Nr. 1-3, 1. April 1999 (1999-04-01), Seiten 8-15, XP002223542 in der Anmeldung erw{hnt *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007050017A1 (fr) * 2005-10-28 2007-05-03 Ge Healthcare Bio-Sciences Ab Support de séparation ayant différentes fonctionnalités
CN102335595A (zh) * 2010-07-20 2012-02-01 华东理工大学 一种新型的具有梯度分布的整体柱固定相的制备方法

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WO2003004536A3 (fr) 2003-03-20
DE10132131A1 (de) 2003-12-04

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