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WO1989011493A1 - Procede de preparation de particules d'agarose - Google Patents

Procede de preparation de particules d'agarose Download PDF

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Publication number
WO1989011493A1
WO1989011493A1 PCT/SE1989/000281 SE8900281W WO8911493A1 WO 1989011493 A1 WO1989011493 A1 WO 1989011493A1 SE 8900281 W SE8900281 W SE 8900281W WO 8911493 A1 WO8911493 A1 WO 8911493A1
Authority
WO
WIPO (PCT)
Prior art keywords
water
polysaccharide
protective colloid
agarose
particles
Prior art date
Application number
PCT/SE1989/000281
Other languages
English (en)
Inventor
Sten Porrvik
Gunnar Mattsson
Original Assignee
Casco Nobel Ab
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 Casco Nobel Ab filed Critical Casco Nobel Ab
Publication of WO1989011493A1 publication Critical patent/WO1989011493A1/fr

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
    • 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
    • B01J20/291—Gel sorbents
    • 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
    • B01J20/24—Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/09—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in organic liquids
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/12—Powdering or granulating
    • C08J3/16—Powdering or granulating by coagulating dispersions
    • 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
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/12—Agar-agar; Derivatives thereof

Definitions

  • agarose gel particles For preparing agarose gel particles, the normal procedure is first to prepare a solution of agarose in water at a temperature of 80-100°C, yielding a viscous a ⁇ ueous solution. This solution is then emulsified at an elevated temperature in an organic phase in the presence of a suitable surfactant. This gives a water-in-oil emulsion with the viscous droplets of agarose-aqueous solution as discontinuous phase. By cooling the hot W/O-emulsion, the agarose molecules crystallise, with the formation of gel particles of a specific pore structure. The pore size can be affected by the content of agarose in the aqueous solution. The higher the agarose content is, the finer are the pores.
  • the pore size is of vital importance since the gel particles are often used for separating biomolecules according to size, so-called molecule sieving. If an aqueous solution of different proteins is supplied to a column containing gel particles, a separation of the proteins according to size will take place in the sense that the smaller molecules will penetrate deeper into the network of the gel particles and, thus, be delayed in the column. The larger protein molecules will therefore leave the chromatographic column first. Protein molecules above a certain maximum size are completely unable to penetrate into the pores and, hence, will pass unimpededly through the column.
  • the emulsifying system makes it possible to prepare gel particles of different average particle size.
  • the present invention relates to a method for preparing particles of polysaccharides, using an inorganic protective colloid which satisfies to a considerable extent the requirements set out above, as appears from the accompanying claims.
  • the emulsion Before isolating the gel particles from the organic phase, the emulsion can be acidified, optionally after adding water, for dissolving the protective colloid.
  • the dry solids content of the gel particles prepared is within the range of from 0.5 to about 50% by weight.
  • Examples of useful inorganic compounds sparingly soluble in water are different sparingly soluble salts of phosphate or polyphosphate, of sulphate, carbonate or silicate. It is also possible to use sparingly soluble oxides or fine-grained minerals. Specific examples of suitable inorganic compounds sparingly soluble in water are calcium phosphate, calcium sulphate, calcium carbonate, iron phosphate, magnesium hydroxide, aluminium oxide, aluminium hydroxide, silicon dioxide (silicic acid), iron hydroxide, barium sulphate, zinc oxide, fine-grained glass powder, bentonite, titanium dioxide etc.
  • the amount of protective colloid should be within the range of from 0.1 to 50%, preferably from 0.3% to 10% based on the amount of the phase of polysaccharide solution.
  • suitable co-stabilisers for hydrophobating the particle surface of the sparingly soluble inorganic compounds are anionic, cationic organic compounds, alcohols, amines or derivatives of ethylene oxide or propylene oxide. Specific examples are carboxylic acids, mono- or diesters of phosphoric acid, alkyl sulphonic acids and quaternary ammonium compounds etc.
  • the particles of the sparingly soluble inorganic compounds may however also be made hydrophobic by bonding organic compounds to the surface of the particles.
  • One example of such a case is treating particles of silicon dioxide with methyl silane to obtain hydrophobic methyl silanol groups on the particle surface.
  • the hydrophobicity of the inorganic protective colloid varies with the type of polysaccharide and inorganic compound, the desired particle size and, primarily, the type of organic phase. By simple tests for a certain system, the amount of co-stabiliser can be easily determined.
  • examples of gel-forming polysaccharides are agar/agarose and dextran.
  • the invention relates to the preparation of agarose particles.
  • W/O-emulsions with the narrow particle size distribution which is characteristic of the system according to the invention if a polysaccharide-aqueous solution having a high polysaccharide content is used in the emulsification.
  • a narrower particle size distribution is generally obtained when using agarose solutions having a lower dry solids content.
  • gel particles having a high dry solids content generally within the range of from 5 to about 30% by weight, and a narrow particle size distribution.
  • a W/O-emulsion having a lower dry solids content preferably 0.5-5% by weight, is first prepared, and water is then removed from the droplets of polysaccharide-aqueous solution at a temperature above the temperature at which the polysaccharide solidifies so as to form a solid gel.
  • This concentration by removing water can be carried out in different ways:
  • This test gave a narrow particle size distribution with 80% by weight in the range of from 40 to 60 ⁇ m. Further tests showed that it was possible to adjust the particle size distribution by the amount of colloid added.
  • An agarose solution was emulsified according to Example 7. At 95°C, an extra 400 g hexanol and 0.75 g lauric acid were added before the sample was cooled to room temperature. After washing the agarose gel particles, the sample was filtered so as to yield a dry cake, and the dry solids content was determined at 9.3%. Similarly, the dry solids content was determined for a sample without any extra hexanol at 4.5%. The density of the aqueous hexanol was determined at 0.830 by means of an aerometer and at

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Colloid Chemistry (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Abstract

Un procédé permet de préparer des particules de polysaccharides gélifiants utiles comme matrices colloïdales de séparation de différentes biomolécules, telles que des protéines ou des acides nucléiques. On prépare les particules colloïdales en dissolvant le polysaccharide dans de l'eau à une température supérieure à sa température de dissolution et en émulsifiant la solution dans un solvant organique de façon à former une émulsion d'eau dans de l'huile, puis en refroidissant l'émulsion jusqu'à ce que le polysaccharide se solidifie et se sépare du solvant. Afin d'obtenir des particules ayant une répartition granulométrique très étroite, on procède à l'émulsification décrite en présence d'un colloïde inorganique hydrophobe de protection.
PCT/SE1989/000281 1988-05-25 1989-05-19 Procede de preparation de particules d'agarose WO1989011493A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE8801952-6 1988-05-25
SE8801952A SE461149B (sv) 1988-05-25 1988-05-25 Saett att framstaella partiklar av gelbildande polysackarider

Publications (1)

Publication Number Publication Date
WO1989011493A1 true WO1989011493A1 (fr) 1989-11-30

Family

ID=20372427

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE1989/000281 WO1989011493A1 (fr) 1988-05-25 1989-05-19 Procede de preparation de particules d'agarose

Country Status (2)

Country Link
SE (1) SE461149B (fr)
WO (1) WO1989011493A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019115A1 (fr) * 1992-03-18 1993-09-30 Pharmacia Lkb Biotechnology Ab Gels polysaccharidiques superporeux
WO2000017257A1 (fr) * 1998-09-23 2000-03-30 Amersham Pharmacia Biotech Ab Production de billes de polysaccharide
WO2006033634A1 (fr) * 2004-09-22 2006-03-30 Ge Healthcare Bio-Sciences Ab Procede de preparation d'une matrice de chromatographie
WO2012156508A1 (fr) * 2011-05-18 2012-11-22 Süd-Chemie AG Procédé pour la préparation de milieux de séparation pour la purification et/ou l'isolement d'enzymes et/ou de protéines
WO2020221762A1 (fr) 2019-04-29 2020-11-05 Cytiva Bioprocess R&D Ab Procédé de fabrication de billes d'agar ou d'agarose
CN114191848A (zh) * 2021-12-06 2022-03-18 武汉瑞法医疗器械有限公司 琼脂糖微球的清洗方法
CN114773497A (zh) * 2022-05-16 2022-07-22 溯玄(上海)生物技术有限公司 一种玫瑰多糖和促进皮肤更新修复及增加皮肤弹性、抗皮肤衰老的用途
WO2023287348A1 (fr) 2021-07-15 2023-01-19 Bio-Works Sweden Ab Procédé de fabrication de billes d'agar ou d'agarose à l'aide d'huile naturelle ou végétale

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB1452896A (en) * 1972-12-21 1976-10-20 Nat Res Dev Packing materials
SE403116B (sv) * 1970-06-25 1978-07-31 Exploaterings Ab Tbf Stabiliserad agarprodukt samt sett for dess stabilisering

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE403116B (sv) * 1970-06-25 1978-07-31 Exploaterings Ab Tbf Stabiliserad agarprodukt samt sett for dess stabilisering
GB1452896A (en) * 1972-12-21 1976-10-20 Nat Res Dev Packing materials

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993019115A1 (fr) * 1992-03-18 1993-09-30 Pharmacia Lkb Biotechnology Ab Gels polysaccharidiques superporeux
US5723601A (en) * 1992-03-18 1998-03-03 Pharmacia Biotech Ab Super porous polysaccharide gels
WO2000017257A1 (fr) * 1998-09-23 2000-03-30 Amersham Pharmacia Biotech Ab Production de billes de polysaccharide
US6590096B1 (en) 1998-09-23 2003-07-08 Amersham Biosciences Ab Process for the production of polysaccharide beads
WO2006033634A1 (fr) * 2004-09-22 2006-03-30 Ge Healthcare Bio-Sciences Ab Procede de preparation d'une matrice de chromatographie
RU2367517C2 (ru) * 2004-09-22 2009-09-20 Джи-И Хелткер Байо-Сайенсиз АБ Способ получения хроматографической матрицы
WO2012156508A1 (fr) * 2011-05-18 2012-11-22 Süd-Chemie AG Procédé pour la préparation de milieux de séparation pour la purification et/ou l'isolement d'enzymes et/ou de protéines
CN113710728A (zh) * 2019-04-29 2021-11-26 思拓凡生物工艺研发有限公司 制造琼脂或琼脂糖珠的方法
WO2020221762A1 (fr) 2019-04-29 2020-11-05 Cytiva Bioprocess R&D Ab Procédé de fabrication de billes d'agar ou d'agarose
JP2022530640A (ja) * 2019-04-29 2022-06-30 サイティバ・バイオプロセス・アールアンドディ・アクチボラグ アガー又はアガロースビーズを製造する方法
CN113710728B (zh) * 2019-04-29 2025-02-11 思拓凡生物工艺研发有限公司 制造琼脂或琼脂糖珠的方法
US12251681B2 (en) 2019-04-29 2025-03-18 Cytiva Bioprocess R&D Ab Method of manufacturing agar or agarose beads
WO2023287348A1 (fr) 2021-07-15 2023-01-19 Bio-Works Sweden Ab Procédé de fabrication de billes d'agar ou d'agarose à l'aide d'huile naturelle ou végétale
KR20240036554A (ko) 2021-07-15 2024-03-20 바이오-웍스 테크놀로지스 에이비 천연 또는 식물성 오일을 사용한 아가 또는 아가로스 비드의 제조 방법
CN114191848A (zh) * 2021-12-06 2022-03-18 武汉瑞法医疗器械有限公司 琼脂糖微球的清洗方法
CN114191848B (zh) * 2021-12-06 2022-12-30 武汉瑞法医疗器械有限公司 琼脂糖微球的清洗方法
CN114773497A (zh) * 2022-05-16 2022-07-22 溯玄(上海)生物技术有限公司 一种玫瑰多糖和促进皮肤更新修复及增加皮肤弹性、抗皮肤衰老的用途

Also Published As

Publication number Publication date
SE461149B (sv) 1990-01-15
SE8801952D0 (sv) 1988-05-25
SE8801952L (fr) 1989-11-26

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