EP1231992A1 - Procede de separation de cellules et de biomolecules par chromatographie a contre-courant - Google Patents

Procede de separation de cellules et de biomolecules par chromatographie a contre-courant

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Publication number
EP1231992A1
EP1231992A1 EP00987272A EP00987272A EP1231992A1 EP 1231992 A1 EP1231992 A1 EP 1231992A1 EP 00987272 A EP00987272 A EP 00987272A EP 00987272 A EP00987272 A EP 00987272A EP 1231992 A1 EP1231992 A1 EP 1231992A1
Authority
EP
European Patent Office
Prior art keywords
separation
modified
biomolecules
cells
unmodified
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Withdrawn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
EP00987272A
Other languages
German (de)
English (en)
Inventor
Renate BLÜTTERS-SAWATZKI
Peter Czermak
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.)
Scinu Tec GmbH
Original Assignee
Scinu Tec 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.)
Filing date
Publication date
Application filed by Scinu Tec GmbH filed Critical Scinu Tec GmbH
Publication of EP1231992A1 publication Critical patent/EP1231992A1/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Withdrawn legal-status Critical Current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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28023—Fibres or filaments
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C3/00—Preservation of milk or milk preparations
    • A23C3/07—Preservation of milk or milk preparations by irradiation, e.g. by microwaves ; by sonic or ultrasonic waves
    • A23C3/076—Preservation of milk or milk preparations by irradiation, e.g. by microwaves ; by sonic or ultrasonic waves by ultraviolet or infrared radiation
    • 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
    • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
    • B01D15/1814—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns recycling of the fraction to be distributed
    • B01D15/1821—Simulated moving beds
    • B01D15/185—Simulated moving beds characterized by the components to be separated
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
    • B01D15/3804—Affinity chromatography
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2215/00—Separating processes involving the treatment of liquids with adsorbents
    • B01D2215/02—Separating processes involving the treatment of liquids with adsorbents with moving adsorbents
    • B01D2215/023—Simulated moving beds

Definitions

  • the invention relates to a method for the separation of cells and biomolecules from mixtures which contain these cells and biomolecules by means of continuous countercurrent chromatography with continuous sample application and removal using solid separation materials.
  • chromatographic processes have already been developed for the purification of biological substances or substance mixtures. These include, for example, HPLC, anion exchange chromatography, gel filtration, affinity chromatography and hydrophobic matrix chromatography.
  • These methods also include those that, for. B. work with affinity chromatography.
  • mild application media are used, from which the substance to be obtained is removed by relatively strong binding to the separation medium.
  • the subsequent selective detachment from the separation medium then always requires the use of a relatively drastic so-called elution medium, with the aid of which the substance sought can then be obtained in a second separate step.
  • elution media are usually very rich in salt, and therefore the substances sought have to be freed from the salt load in a very complex manner.
  • DE 1 9938394 A1 and DE 3930947 A1 describe affinity chromatography methods which use absorption on surfaces.
  • the molecules to be separated such as antibodies, are covalently bound to the separation surfaces by means of strong bonds. Due to the strong connection to such surfaces, the selective detachment of the corresponding substances can only take place by using specific elution media, the substance previously bound to the support being detached by this elution medium in a second separate step. Thus, such methods cannot be used for continuous Chromatography methods, such as the countercurrent method described above, can be used.
  • Continuous chromatographic processes have therefore already been developed in which the fluid to be chromatographed or a multicomponent mixture is fed continuously and the separated fractions are also continuously removed.
  • Some of these are so-called countercurrent processes.
  • a representative of such a countercurrent process is, for example, the so-called simulated moving bed process (SMB).
  • SMB simulated moving bed process
  • SMB simulated moving bed process
  • TMB True Moving Bed
  • the so-called True Moving Bed (TMB) process also belongs to these continuous countercurrent processes.
  • the time-dependent chromatographic separation of the column chromatography is converted into a spatially resolved separation.
  • This known countercurrent process mainly uses porous, particulate separation media.
  • porous materials have always been used, as described, for example, in WO 9803242.
  • the delaying effect that causes the separation of the biomolecules is a different flow of the molecules through the pores or past the pores, depending on the size of the molecule.
  • Interactions are examples of such weak interactions.
  • Weak interactions are understood to mean those bonds, which are typically typical of biomolecules, that are not covalent in nature and are relatively unstable, such as e.g. B. the already known hydrogen bonds, hydrophobic and electrostatic bonds and van der Waals forces.
  • the description of such weak interactions, as are known for many interactions between biomolecules, can be thermodynamically with the aid of the "heat capacity difference" ( ⁇ C P ), as described in Cooper (A.
  • the object of the present invention is accordingly to provide a method for separating cells and biomolecules from mixtures, in particular from complex mixtures, by means of countercurrent chromatography with continuous sample application and removal using solid non-porous separation materials, with which in the mixtures contained cells and biomolecules can be separated or separated, especially for production purposes and on an industrial scale.
  • a core element of the method according to the invention is that the weak interactions described in the context of the present documents are used.
  • the above-discussed dissociation constant K d which according to the invention is greater than or equal to 10 ⁇ 5 M, is used to define these weak interactions. This definition is therefore used in the context of the invention to characterize weak interactions.
  • the process according to the invention is a continuous process or a continuous process which works according to the countercurrent process, with only a single liquid process Separation medium is used.
  • Non-porous separating materials are understood to mean those materials that are already partially used in materials such as nylon, polystyrene, silicate glass (AL Plant et al., Immobilization of binding proteins on nonporous supports.
  • Quartz fibers P. Wikstrom. Larsson PO, Affinity fiber - a new support for rapid enzyme purification by high-performance liquid affinity chromatography, J Chromatogr 1987, 388: 123-34.
  • the applications described also did not relate to separations in which so-called weak interactions were used as the separation principle.
  • the already known retarding effect due to diffusion processes which can be observed with porous materials, is used, but a completely different principle.
  • a delaying effect is necessary in the method according to the invention for separating the molecules.
  • this is achieved by the weak interactions described in more detail above, which the molecules to be separated enter into with the surfaces of the non-porous separation materials.
  • the surfaces can either be modified or not modified.
  • the appropriate selection of the surfaces or their modification ensures that the corresponding molecules to be separated are retained by molecular adhesion effects on the corresponding surfaces, as a result of which the molecules are separated.
  • the degree of separation can be affected by the density of the corresponding molecular attachment sites.
  • the process according to the invention which exploits the weak interactions described, has the advantage that the biomolecules can be separated without the use of drastic measures, for example high salt concentrations. High pressure differences can thus be avoided in the method according to the invention, so that a separation of cells from biological matrices can also be achieved.
  • the separation takes place - as described - basically on surfaces that are formed on non-porous matrices or porous membranes.
  • the method according to the invention can be used in particular for the industrial separation of biomolecules and cells.
  • biomolecules proteins, carbohydrates, nucleic acids, glycoproteins, glycolipids, vitamins, enzymes, lipids and bioactive substances can be used as biomolecules.
  • hormones and phytoestrogens are suitable as bioactive substances.
  • the cells used are preferably those from plant, animal or unicellular cell sources or from peripheral blood. It may be necessary to use different separation matrices for certain molecules of these substance classes.
  • the separation materials are selected in such a way that the weak interactions are sufficient to separate the substances from one another without them permanently adhering to the surfaces.
  • the density of the release-active surface components or adhesion sites is preferably set such that no additions of salts are required for the separation.
  • the method according to the invention is primarily intended for the separation of peripheral stem cells from the blood of human donors.
  • the blood is preferably introduced directly into a device for carrying out the method according to the invention, which is described in more detail below, while the cells in question are retarded on the corresponding surfaces in such a way that the stem cells can be removed continuously from the device, while the other blood components in the can be removed substantially unchanged at other points in the device.
  • cells which can be obtained from fermentations are preferably used as cells. It can be act plant, animal or unicellular organisms. According to the invention, primarily biomolecules and thus those molecules that come from biological sources are taken into account for the industrial separation processes. These biomolecules represent important starting products for the pharmaceutical industry and for the food industry. According to the invention, animal milk and dairy products, fermentation supernatants, molasses, plant extracts, residues from plant processing, blood products, wash water are therefore preferably used as the liquid to be chromatographed or as a multi-component mixture
  • the separating materials used can have unmodified and also modified surfaces.
  • the modification of surfaces is known per se and is used for the separation using porous, particulate separation media or using porous membranes.
  • porous, particulate separation media or using porous membranes In contrast to
  • Biological polymers are preferably used as separation materials with an unmodified surface. These preferably include celluloses, chitosans, dextrans,
  • Modified biological polymers are also preferably used as separating materials. These preferably include modified celluloses such as cellulose acetates, carboxymethyicellulose,
  • PVC polyvinyl chloride
  • polyacrylonitrile polyaramid
  • polystyrenes including styrene / divinylbenzene polymers
  • polyester polypropylene
  • teflone polypropylene
  • PTFE teflone
  • FEP FEP
  • PFA polyvinyl alcohols
  • polyvinylidene fluoride polyvinyl difluoride
  • polyolefins methacrylic polymers
  • polybenzimidazole polyfuranethylene
  • polyphenols polyphenols
  • Polybutadienes Polybutadienes, polybenzimidazolone, polyether / amide, polyether / urea, polyether viscose, polydimethylsiloxane, silicone, siloxanes, polycarbonate, polyalkylsulfone, polyisoprene, polyethylene terephthalate and mixtures of two or more of these plastics.
  • modified and unmodified inorganic matrices are preferably used, these being, in particular, biocompatible materials such as carbon / carbon fiber composites, ceramics (TiO 2 , Al 2 O 3 , ZrSiO 4 , ZrO 2 ), silicates, glasses and other matrices (e.g. Fabrics made of metal wire or glass fiber with or without surface modification), which are typical for chemical apparatus construction. It must also be taken into account here that the biomolecules and cells to be separated do not penetrate through or into the surfaces in order to avoid the possibility of uncontrolled absorption and unwanted pressure drops.
  • the modification of the separation materials (more precisely the surfaces thereof) can be carried out using methods known per se.
  • the possible methods of modification preferably include chemical linking (cross-linking).
  • Groups present on the polymers for example hydroxyl, carboxy, carboxyl, amino, amide, aryl, sulfhydryl, alkyl, methoxy and ethoxy radicals or groups, are activated by methods known per se as described, for example, in: E. Müller, E. Klein “Membranes Modified for Biochromatography” in: Bioseparation and Bioprocessing. Vol 1 (Editor: G. Subramanian), Wiley-VCH, Weinheim 1 997.
  • the required ligands are covalently bound to the surfaces, and the activation methods which can be used according to the invention include, for example, methods in which the following activation reagents are used: cyanogen bromide, tosyl chloride, N-hydroxy succinimide, carbonyl-di-imidazole, carbodiimides, 1, 4 -Butanediol-diglycidol ether, glutaraldehyde and 2-fluoro-1-methylpyridine If desired, so-called spacers can be introduced as attachment sites or binding sites for the weak interaction Can then attach to the activated surfaces preferably the following biomolecules are bound: polymeric carbohydrates (e.g.
  • chitosans and dextrans glycoproteins, glycopeptides, glycans, lectins, polymerized amino acids (e.g. poly-L-leucine), peptides, proteins, nucleotides and biomolecule-binding dyes.
  • the geometry of the surfaces of the separating materials on which the separation takes place can be designed in a very varied manner.
  • the separating materials are washed or flushed by the liquid to be chromatographed.
  • So z. B smooth surfaces in the smallest possible distances in the form of narrow channels or tubes.
  • Flexible hoses with small diameters can also be used.
  • spacers in the form of threads nets or precision fabrics are preferably inserted between the surfaces. These spacers can be made of the same material as the surfaces of the separating materials.
  • tubes, hoses or winding units made of an inert material can also be used, in the lumens of which there are threads, nets or wick-like fabrics, the surface of which is as previously described.
  • Irregular filter shapes such as molten metal filters (e.g. from Drache, Dietz, FRG), which have a large surface area and are made of ceramic, chemically and thermally highly stressable material can also be used. The liquid flows run past these surfaces and do not penetrate them. Since the liquid flows do not encounter large resistances, large pressure drops can be avoided. This means that pumps and valves can also be used that are sufficient for medium pressure conditions and also less expensive than is the case for high-pressure equipment.
  • the device for carrying out the method according to the invention is provided with several so-called separation units, in which there are separating materials.
  • the separation units are at least chromatographically essentially the same and preferably essentially identical.
  • the separating materials used are preferably approximately the same in terms of shape, size and type and preferably essentially identical.
  • a plurality of separation units are preferably connected in series in terms of flow and combined to form separation zones.
  • valve units which are controlled by a computer and whose position is changed using a mathematical model, so that the weak separation effect based on the interactions is optimized in such a way that the separation of the substances can be done continuously.
  • the material flows are carried in and out with the aid of several and typically four pumps.
  • the flow and circulatory system which is maintained by another pump, is designed like SMB chromatography. There are therefore several separation zones, each of which is assigned a certain number of separation units and which are coupled to the corresponding lines by valve circuits.
  • the separation method according to the invention which uses the weak interaction mentioned and in which the separation takes place on non-porous surfaces, can also be carried out as computer-controlled countercurrent chromatography by separation on surfaces or in English “Computer controlled counter current chromatography using separation on surfaces” or abbreviated “ C-5-S-2 ". Only the term “C-5-S-2" is used below.
  • FIG. 1 shows the structure of a device for performing the method according to the invention in a highly simplified and very schematic manner, numerous details being omitted in order to be able to illustrate the principle of the method according to the invention more clearly and simply.
  • the structure and the control shown there are - as shown - of a known type. In this regard, reference is made to the publications mentioned above.
  • the individual separation units A, B, C, D, E, F, G and H of the device shown in Fig. 1 are connected in series in terms of flow. Between these individual separation units there are inflows and outflows 1, 2, 3, 4, 5, 6, 7 and 8 to a large valve unit 10.
  • the inflows and outflows 1 to 8 each have an inflow and an outflow; for the sake of simplicity, the two lines are only shown with a line 1 to 8 in the figure.
  • the valve unit 10 is computer-controlled and equipped with two inflows 21 and 22 and two outlets 23 and 24, with one inflow 21 for each Separation medium and an inlet 22 for the mixture to be separated and an outlet 23 for component 1 and an outlet 24 for component 2 are present.
  • two separation units A, B; C, D; E, F and G, H connected to separation zones AB, CD, EF and GH.
  • the inflows and outflows are switched in such a way by the valve arrangement that an inflow and outflow always alternate.
  • the first separation zone AB is thus between the inflow of the separation medium and the outflow of component 2 (extract).
  • the second separation zone CD is located between the outflow of component 2 (extract) and the inflow of the mixture to be separated.
  • the third zone EF is here between the supply of the mixture to be separated and the outflow of component 1 (raffinate).
  • the fourth zone GH is located between the outflow of component 1 (raffinate) and the inflow of the separation medium.
  • two separation units are connected to a separation zone.
  • this type of circuit is only one embodiment.
  • the separation units can also be used alone or connected in groups of three, four, etc.
  • the central fourfold distributor valve unit 1 0 is controlled with the aid of a computer program and can have different mechanical structures.
  • it can be designed as a rotary multiple valve or can be constructed from several triple or single valves. It is only important that the valves are opened and closed in such a way that the separation zones AB, CD, EF and GH are switched on in the direction of flow of the mobile or liquid phase in such a way that a quasi counterflow to the solid phase is generated.
  • the pumps are preferably controlled by a computer program in such a way that the flow rates in the separation zones are regulated in such a way that only the pure components 1 and 2 are discharged. It is a complex controlled process in which the suitable parameters are expediently determined by computer simulation.
  • the separation units are manufactured as follows:
  • Teflon tubing 8 mm in diameter is terminated at a length of 40 centimeters at the ends with stainless steel nets, and the tapered ends are each connected to pumps or valves with thinner tubing.
  • a wick-like nylon fabric is inserted between the ends as a separating material, on which the glycan sialyl-Lewis A is coupled as an adhesion phase by activation with cyanuric chloride and dicyclohexylcarbodiimide.
  • the group density was chosen so that less than 0.1 ⁇ mol of the ligand is bound per gram of nylon.
  • Such a piece of Teflon tubing represents a separation unit. A total of 8 separation units are connected in series, the separation zones each consisting of 2,2,3,1 units.
  • Separation zone 1 (separation medium / stem cell extract) consists of 2 units.
  • Separation zone 2 (stem cell extract / blood flow) consists of 2 units.
  • Separation zone 3 (blood inflow / blood outflow) consists of 3 units.
  • Separation zone 4 (blood drain / separation medium) consists of 1 unit. Isotonic saline is used as the separation medium. The entire apparatus is heated to 37 degrees Celsius.
  • Example 2 In this way it is possible to obtain around 10 million stem cells from the peripheral blood of a donor within 2 hours.
  • Example 2
  • Lactoferrin is separated from cow whey using the C-5-S-2 process.
  • the separation units are manufactured as follows. Spacers (nets made of woven cellulose acetate fibers of 0.5 mm diameter) are inserted into tubes of 50 mm diameter made of cellulose acetate and wound flat on cores in lengths of 3 m. The tubing is non-porous and the ends are each sealed and tapered with plastic frit so that the appropriate inflow and outflow tubing can be connected to the units.
  • the cellulose acetate surfaces are activated with the aid of 1.1 carbonylimidazole and chitosans are covalently bound to them.
  • the group density is approximately 1.0 mmol per gram of cellulose acetate.
  • Separation zone 1 (separation medium / lactoferrin outflow) is 1 unit, separation zone 2 (lactoferrin outflow / inflow whey) 4 units, separation zone 3 (inflow whey / outflow residual whey) 4 units and separation zone 4 (outflow residual whey / separation medium) ) 1 unit in size.

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Food Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Toxicology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Sustainable Development (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Abstract

L'invention concerne un procédé de séparation de cellules et de biomolécules à partir de mélanges comprenant ces cellules et ces biomolécules. A cet effet, on utilise la chromatographie à contre-courant avec alimentation et prélèvement permanents d'échantillons et utilisation de matériaux de séparation solides et non poreux. Le procédé est caractérisé en ce qu'un seul matériau de séparation est utilisé, de manière à éviter tout changement de ce matériau, en ce que les matériaux de séparation non poreux utilisés ont une surface modifiée ou non modifiée, et en ce que, pour la séparation des cellules et des biomolécules à séparer, des </= interactions faibles >/= sont mises en oeuvre et résultent de la mise en contact des cellules et des biomolécules à séparer avec la surface modifiée ou non modifiée. Par interactions faibles, il faut comprendre celles dont les liaisons ont une constante de dissociation égale ou supérieure à 10<-5> M (Kd >/= 10<-5> M).
EP00987272A 1999-11-20 2000-11-20 Procede de separation de cellules et de biomolecules par chromatographie a contre-courant Withdrawn EP1231992A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19956010 1999-11-20
DE19956010 1999-11-20
PCT/EP2000/011520 WO2001037959A1 (fr) 1999-11-20 2000-11-20 Procede de separation de cellules et de biomolecules par chromatographie a contre-courant

Publications (1)

Publication Number Publication Date
EP1231992A1 true EP1231992A1 (fr) 2002-08-21

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EP00987272A Withdrawn EP1231992A1 (fr) 1999-11-20 2000-11-20 Procede de separation de cellules et de biomolecules par chromatographie a contre-courant

Country Status (3)

Country Link
EP (1) EP1231992A1 (fr)
AU (1) AU2358101A (fr)
WO (1) WO2001037959A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ539297A (en) * 2002-09-13 2008-04-30 Biogen Idec Inc Method of purifying polypeptides by simulated moving bed chromatography
DE102006028129B4 (de) * 2006-06-15 2008-07-03 Nikolaus Stephan Pfeiffer Kontinuierliche Gegenstom Chromatographie Anlage
CN101678244B (zh) * 2007-04-17 2013-06-19 深度控股有限公司 用于连续膜吸附的方法和装置
CN103736296B (zh) * 2013-12-30 2015-12-30 浙江大学 一种制备丹参酮化合物的双柱循环分离系统及其方法

Family Cites Families (7)

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Publication number Priority date Publication date Assignee Title
SE468653B (sv) * 1987-03-06 1993-02-22 Perstorp Biolytica Adsorbent foer isokratisk affinitetskromatografi
DE3900272A1 (de) * 1989-01-07 1990-07-12 Mueller Schulte Detlef Dr Synthetisches polymeres traegermaterial fuer chromatographische trennverfahren, verfahren zu seiner herstellung und verwendung
GB2274843B (en) * 1993-02-09 1997-02-26 Agricultural & Food Res Continuous separation and purification of materials
US5482631A (en) * 1994-10-06 1996-01-09 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Separation of inositols from sugars and sugar alcohols
DE19629208A1 (de) * 1995-01-20 1998-01-22 Merck Patent Gmbh Verwendung von modifizierten Membranen für "Simulated Moving Bed" Trennverfahren
US5645729A (en) * 1995-02-13 1997-07-08 Uop Separations by simulated moving bed chromatography operating at low k' values using weakly interacting adsorbents as the stationary phase
US5626762A (en) * 1995-02-13 1997-05-06 Uop Separations by simulated moving bed chromatography operating at low K' values using weakly interating adsorbents as the stationary phase

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0137959A1 *

Also Published As

Publication number Publication date
AU2358101A (en) 2001-06-04
WO2001037959A1 (fr) 2001-05-31

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