EP1183531A1 - Annularchromatograph - Google Patents
AnnularchromatographInfo
- Publication number
- EP1183531A1 EP1183531A1 EP98958716A EP98958716A EP1183531A1 EP 1183531 A1 EP1183531 A1 EP 1183531A1 EP 98958716 A EP98958716 A EP 98958716A EP 98958716 A EP98958716 A EP 98958716A EP 1183531 A1 EP1183531 A1 EP 1183531A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- gels
- zone
- annular
- annular chromatograph
- chromatograph according
- 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- Withdrawn
Links
Classifications
-
- 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
- B01D15/1892—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns the sorbent material moving as a whole, e.g. continuous annular chromatography, true moving beds
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/58—Conditioning of the sorbent material or stationary liquid the sorbent moving as a whole
-
- 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
- B01D2215/022—Physically moving the adsorbent as a whole, e.g. belts, discs or sheets
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/58—Conditioning of the sorbent material or stationary liquid the sorbent moving as a whole
- G01N2030/587—Continuous annular chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
- G01N30/463—Flow patterns using more than one column with serial coupling of separation columns for multidimensional chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
Definitions
- the present invention relates to a circular chromatograph with a feed in the form of a particle bed.
- Annular chromatography has been a variant of preparative chromatographic separations that has been recognized for a number of years and is increasingly being used. Annular chromatography is preferably used when large amounts of substance mixtures have to be separated, since this type of chromatography can be operated continuously, and with a relatively high degree of resolution.
- P-CAC preparative continuous annular chromatography
- Such procedures are known and widely used in the prior art (see e.g. EP-A-371 .648).
- annular chromatography In addition to the advantage of the high throughput of annular chromatography, this is also characterized by high resolution and specificity. Nevertheless, for various specific separation problems it is necessary to follow up further annular chromatography steps - for example using different separation gels than in the first step - in order to achieve the desired degree of separation. On the one hand, this is relatively easy to achieve in comparison to conventional column chromatography, since in the case of annular chromatographic separations it is known at which point on the circumference the desired product or products emerge; on the other hand, further annular chromatographs are required for this, which of course greatly increases the outlay in terms of equipment and therefore money.
- the aim of the present invention is therefore to provide a device for annular chromatography, by means of which the resolution and specificity of this chromatographic separation process can be increased efficiently without excessively increasing the outlay on equipment and thus the process costs.
- the invention achieves this aim by means of an annular chromatograph with a feed in the form of a particle bed, which is characterized in that the particle bed has at least two zones arranged one above the other, which contain different bed material and are preferably separated from one another by at least one separating layer.
- At least one of the zones can be adapted as an electrophoresis zone by providing electrical connections, preferably in the form of sliding contacts, as a result of which electrophoretic separations can also be carried out in the advantageous manner described above.
- This electrophoresis zone can preferably be delimited by at least one electrically non-conductive separating layer to the other zone (s) in order to increase the efficiency of the electrophoresis.
- the bed material for the at least two zones can generally consist of anion exchange resins, cation exchange resins, exclusion gels, gel permeation gels, affinity gels, hydrophobchromatography (HIC) gels, displacement (displacement) resins, reversed phase (reversed phase) gels, Electrophoresis gels and other separation media used in practice, which offers a variety of possibilities for combined separations, as described in more detail below. If one of the zones in the chromatograph according to the invention is an electrophoresis zone, an electrophoresis gel is of course chosen as the particle bed for this zone.
- the separating layer or separating layers are preferably selected from membranes, non-porous, inert particle material, in particular glass beads, and - especially for any electrophoresis zones - from electrically non-conductive material.
- the uppermost zone of the particle bed is covered with a cover layer and / or underlaid with a base layer, the cover and base layers preferably being made of the same material as the separating layer (s).
- a chromatograph according to the invention particularly preferably comprises at least one electrophoresis zone or at least one exclusion gel zone and at least one adsorber resin zone, in particular at least one adsorber resin zone containing an ion exchange resin, which, for example, proteins (one after the other according to their size and their charge), as well as numerous biotechnological process products such as eg enzymes, h-EGF ("human epidermal growth factor”) and immunoglobulins, separated and cleaned extremely selectively and with high resolution.
- a tempering (ie heating or cooling) jacket can also be provided on the inner and / or outer circumference of the annular separating column (11) in order to be able to set the optimum temperature for each separation.
- FIGS. 1 a) and 1 b) are schematic representations of an annular chromatograph with 2 or 3 separation zones that can be used according to the present invention
- FIG. 2a) is a schematic
- FIG. 2b) is a schematic enlargement of a detail of the dashed area from FIG. 2a).
- FIG. 2b) is a partial view of a section through a further annular chromatograph according to the invention with an electrophoresis zone.
- 1 a) and 1 b) two variants of the annular chromatograph of the present invention are shown schematically.
- 1 a) shows an embodiment with two, in FIG. 1 b) one with three separation zones of the particle bed.
- zones 1 and 2, or 1, 2 and 3 preferably consist of release resins selected from e.g. Anion exchange resins, cation exchange resins, exclusion gels, affinity gels, hydrophobchromatography (HIC) gels, displacement (displacement) resins, electrophoresis gels, and optionally from other stationary phases customary in the field, such as e.g. Silica gel, aluminum oxide, etc.
- release resins selected from e.g. Anion exchange resins, cation exchange resins, exclusion gels, affinity gels, hydrophobchromatography (HIC) gels, displacement (displacement) resins, electrophoresis gels, and optionally from other stationary phases customary in the field, such as e.g. Silica gel, aluminum oxide,
- the invention comprises the following combinations of two stationary phases:
- Ion exchange chromatography e.g. Mono S (Pharmacia) or Toyopearl DEAE 650S (TosoHaas);
- Reversed phase chromatography e.g. Amberchrom CG-161 cd (Rohm & Haas) or Sephasil C8 (Pharmacia);
- Size exclusion and gel permeation chromatography e.g. Sephadex G1 5 (Pharmacia) or Toyopearl HW 40F (TosoHaas);
- Affinity chromatography e.g. Toyopearl AF Tresyl-650M (TosoHaas) or EAH Sepharose-4B (Pharmacia);
- Adsorption chromatography e.g. Amberlite XAD (Rohm & Haas) or Purolite MN200 (Purolite).
- two cation exchange gels or two exclusion gels can also be arranged one above the other (eg Sephadex G 15 via Toyopearl HW 40F or the like).
- annular chromatographs with a particle bed comprising at least one exclusion gel zone and at least one adsorber resin zone, in particular at least one adsorber resin zone containing an ion exchange resin, and annular chromatographs with at least one electrophoresis gel in one of the zones.
- the two or three gels are introduced one after the other into the annular gap of an annular column 11 made of material which is inert to the components of the separation solutions, preferably glass, each of the gel zones being covered with a separation layer 5 before the next is applied thereon to prevent mixing of the different zone materials.
- the top of the particle bed forms a cover layer 8.
- a base layer 9 is additionally shown, which serves (in addition to a porous base plate, not shown), for example frit, membrane disc, etc. to prevent particulate matter from escaping at the bottom of the column.
- the material for the separating, covering and base layers 5, 8, 9 is selected from membranes and non-porous particle material which is inert to all components of the respective separating solutions and can be the same or different for all three layers, but it is special must not be electrically conductive for electrophoretic separations. Glass beads are preferred according to the invention, since they are inert and easy to apply in practically all common applications.
- the loading takes place in the order 9-2-5-1 -8, in FIG. 1 b) in the order 3-5-2-5-1-8.
- the other components of the annular chromatograph can be designed in a conventional manner. So the column 1 1 (driven by a motor, not shown) rotatably mounted on an axis 12 and is fed via connecting lines 1 3 for feed and eluents and a distributor head 14 fixedly mounted on the axis, the feed channels 15 of the distributor head 14 preferably immersing in the cover layer 8 in order to ensure an even task.
- the channels can have the usual designs, ie single, multiple or slot nozzles or the like, but curved slot nozzles of different widths which are adapted to the column circumference are preferred for the invention, in order to be able to match the feed and eluent flows as closely as possible.
- outlet channels 16 are provided for collecting the eluates. These outlets 16 can either be connected to the column 11 (i.e. they rotate with it about axis 12) or they can be fixed to the axis 12 and e.g. are in contact with the column rotating relative to it via a slip ring, the latter embodiment being preferred.
- FIG. 2a shows a partial view of an embodiment of an annular chromatograph according to the invention with an electrophoresis separation step, namely the electrophoresis separation zone 4, which leads to another zone above and below (not shown) by means of a separation layer 5 made of electrically non-conductive material is limited.
- Slip contacts are provided both at the upper and at the lower end of the electrophoresis zone 4, preferably directly adjacent to the respective separation layer (s), each consisting of a conductor ring 6 mounted on the outside of the axis 12, each of which has a network connection 10 is supplied with current, as well as from an annular current collector 7 provided on the inside of the column 1 1, which lies closely against the conductor ring 6 and is in electrical contact therewith.
- Example 1 Cation exchanger - exclusion gel: ion exchange-size exclusion chromatography
- the platinum group metals rhodium, palladium, platinum and iridium can be continuously separated from one another by means of preparative annular chromatography. Common metals present at the same time, such as iron, copper, nickel or cobalt, can also be continuously separated from the precious metals.
- the precious metal solution is usually in concentrated hydrochloric acid. Due to the high chloride concentration, the noble metals are all present as anionic chloro complexes.
- Base metals have the property of binding to a cation exchanger under certain conditions, while the noble metals as anionic complexes pass through the cation exchanger without retention. This property can be used to make the base in the first step Separate accompanying metals from the precious metals and separate the precious metals from one another in a second process step. With the P-CAC system of the present invention, this is possible in the simplest apparatus design.
- exclusion gel 2 e.g. Sephadex, Toyopearl or Biogel
- d 1 50-240 ⁇ m
- a cation exchanger 1 e.g. Dowex, Purolite, Amberlite
- another glass bead layer 8 are layered over this glass bead layer 5.
- the feed solution is pumped into the annular column at the radial position 0 °.
- the feed channel 15 is immersed in the upper glass bead layer 8.
- the main eluent (top eluent) is 0.5-1 m Hcl.
- the noble metals run through the cation exchange layer 1 without interaction and are then separated from one another in the exclusion gel layer 2.
- the individual fractions of rhodium (Rh), palladium (Pd), platinum (Pt) and iridium (Ir) can be collected separately.
- the base metals adsorb to the cation exchanger and are stripped at the radial position where the last precious metal elutes (Ir) with a step eluent (2 m or higher HCI).
- the 2 m HCI desorbs the base metals from the cation exchanger and drives them to the exclusion gel layer. There the base metals have no interaction with the stationary phase. The base metals can now be obtained as a common fraction.
- a 26 cm high layer of Sephadex G-15 was covered with 5 cm glass beads.
- a 5 cm thick layer of Dowex 50-W X8 was applied to this layer, which in turn was overlaid with a layer of glass beads.
- the solution was eluted with 0.5 m HCl.
- the noble metals passed through the cation exchange layer 1 without interaction and separated on the Sephadex layer 2.
- Four differently colored bands formed - Rh band (red), Pd band (brown), Pt band (yellow) and Ir band (brown).
- the step eluent (2 m HCI) was stripped for the base metals.
- the step eluent desorbed the base metals from the cation exchanger, and a greenish-yellow band formed that runs through the column without interaction.
- the base metals could be obtained as a common fraction after the iridium fraction.
- Ru and Os are also contained in the separation mixture, Ru elutes between Rh and Pd and Os according to Ir (see Example 2).
- Example 2 Adsorption resin - exclusion gel adsorption Z size exclusion chromatography
- the exclusion gel is filled into the P-CAC (at least up to half the height), a separation layer (glass beads or membrane) is placed over the exclusion gel, the adsorption resin (Amberlite XAD7 or Purolite MN 200) is filled over this layer.
- a separation layer glass beads or membrane
- the adsorption resin Amberlite XAD7 or Purolite MN 200
- a precious metal solution containing all the platinum group metals (but at least two of them) and gold is used as the feed for the P-CAC.
- Gold is from the adsorber resin selectively adsorbed, while the other platinum group metals run unretarded through the adsorbent layer and are separated on the exclusion gel. After elution of the last component, the gold is stripped from the adsorber resin using a step eluent.
- the anion exchanger is filled at least halfway up the P-CAC.
- a separation layer is filled over the anion exchanger (membrane or glass beads of the appropriate size), and over the separation layer the cation exchanger up to the maximum filling level.
- a metal solution containing rhodium and iridium as well as non-precious metals such as iron, nickel, cobalt or copper is pumped into the annular column as a feed.
- the base metals adsorb on the cation exchanger, while rhodium and iridium pass through to the anion exchanger.
- Ir (IV) adsorbs on the anion exchanger and rhodium runs through.
- step eluent 1 After all rhodium is eluted, turn on this angular position with the step eluent 1 elutes the base metals. After this elution, the reduction from Ir (IV) to Ir (III) takes place with the step eluent 2.
- Example 4 Anion exchanger - exclusion gel ion exchange-size exclusion chromatography
- the P-CAC is filled with an exclusion gel at least up to half the fill level, then overlaid with a separating layer, which is in turn overlaid with an anion exchanger.
- a precious metal solution which in addition to gold as a cyanide complex also contains the platinum group metals, is applied to the ring gap as a feed solution.
- the gold cyanide binds as an anionic complex to the anion exchanger, the remaining platinum group metals pass through the anion exchanger without interaction and are separated on the exclusion gel.
- a strip solution for the desorption of the gold is introduced into the annular gap.
- anion exchangers and exclusion gels in biotechnology is also conceivable; eg for the production and purification of monoclonal antibodies.
- the anion exchanger can be used to remove pyrogens, nucleic acids and proteases; the salts of the buffer solution can then be removed in the exclusion gel.
- a P-CAC column is filled with an exclusion gel to at least 50% of the fill level, overlaid with a separating layer and loaded with superlig gels up to the final fill level (superlig gels are gels with crown ether as a functional group; trademark of IBC Advanced Technology) .
- superlig gels are gels with crown ether as a functional group; trademark of IBC Advanced Technology) .
- a precious metal solution containing a high proportion of base metals is applied as a feed to the ring gap of the P-CAC.
- the base metals have no affinity for the Superlig gel and therefore elute through the entire pillar and can be collected as a common fraction.
- the precious metals are stripped from the Superlig gel and separate on the exclusion gel in the order Rh - Pd - Pt - Ir.
- the anion exchanger as the upper layer serves to pre-clean the cellulase; is eluted with a Tris.HCI buffer.
- the fraction obtained, in which the cellulase is located, is then further fractionated in a cation exchanger as under layer (Pharmacia Mono S) with an acetate buffer as the eluent.
- a very high resolution can be achieved with the cation exchanger, so that the cellulase can be obtained in a pure state.
- Example 7 Cation exchanger - exclusion gel ion exchange-size exclusion chromatography
- a preliminary cleaning and concentration of the antibody is achieved with the cation exchanger as the upper phase (Pharmacia S Sepharose High Performance).
- MES with NaCI is used as a buffer.
- the pool (fraction which also contains the antibody) is then freed of dimeric oligomers and of transferrin via an exclusion gel (Superdex 200) as the lower phase with a sterile sodium chloride buffer.
- Example 8 Anion exchanger - exclusion gel ion exchange-size exclusion chromatography
- the anion exchanger (Pharmacia Q - Sepharose) as the upper phase serves for the pre-cleaning (cleaning based on the charge) of the growth factor
- the exclusion gel serves for the final cleaning of the growth factor.
- Example 9 Hydrophobchromatography (HIC) gel exclusion gel
- the upper stationary phase (HIC gel, Pharmacia Phenyl Sepharose CL) is used for the initial cleaning and concentration of the prolactin (concentration by a factor of 7).
- the prolactin is finally purified on an exclusion gel (e.g. Pharmacia Sephadex G 100).
- Protein pool can be obtained that contains proteins of the same charge, including the macroglobulin.
- the macroglobulin is in the second layer, the HIC gel
- the anion exchanger e.g. Pharmacia DEAE Sepharose
- the fermentation broth is separated after loading.
- the pool in which the transcriptase is located passes through the second layer, the HIC gel (e.g. Pharmacia Phenyl Sepharose High Performance). Due to the different hydrophobicity after this layer, the transcriptase is obtained in pure form.
- Example 12 Cation exchanger - anion exchanger, with one of the stationary phases in the displacement (displacement) mode ion exchange-Z-displacement chromatography protein separation
- An anion exchanger eg Pharmacia Mono Q
- An inert layer for example glass beads or a membrane, is layered on this anion exchanger and a cation exchanger is filled in (for example Pharmacia Mono S).
- a mixture of different proteins is separated on the cation exchanger due to different charge, the separated fractions are concentrated on the anion exchanger using a displacement reagent or displacer (eg Nalcolyte). Separation and simultaneous concentration can thus be combined.
- a displacement reagent or displacer eg Nalcolyte
- Example 1 3 Exclusion gel - electrophoresis gel
- the electrophoresis gel forms the lower layer in the P-CAC, which is covered with an exclusion gel. If proteins of different charge and different pI values are now applied to the exclusion gel, they separate due to their size. The electrophoretic layer then takes over the additional separation of the proteins based on their charge.
- Example 14 Exclusion gel - cation exchanger, with the cation exchanger in displacement mode, size exclusion zone exchange chromatography
- an exclusion gel eg Pharmacia Sephadex G25
- proteins eg albumins
- a feed solution which contains the amino acids L-glutamic acid, L-valine and L-leucine. Separation in the exclusion gel gives a fraction containing the proteins and a fraction containing the amino acids.
- the amino acids are separated on a cation exchanger (eg Dowex 50 W-X8) with a displacer (eg 0.1 N NaOH) and concentrated at the same time.
- the amino acids are listed in the order glutamine Acid - valine - leucine obtained.
- the displacer must be followed by a regeneration solution (eg dilute H 2 S0 4 ) in order to bring the gel bed into its original state.
- displacement elution is also not limited to use in cation exchange.
- the present invention provides a new annular chromatograph with a particle bed consisting of several different zones, with which several types of chromatographic separations can be carried out continuously, in one step and thus more quickly and cost-effectively than in the prior art was previously possible.
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AT203097 | 1997-12-01 | ||
AT203097A AT405025B (de) | 1997-12-01 | 1997-12-01 | Annularchromatograph |
PCT/AT1998/000290 WO1999028740A1 (de) | 1997-12-01 | 1998-11-30 | Annularchromatograph |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1183531A1 true EP1183531A1 (de) | 2002-03-06 |
Family
ID=3526306
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP98958716A Withdrawn EP1183531A1 (de) | 1997-12-01 | 1998-11-30 | Annularchromatograph |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP1183531A1 (ru) |
AT (1) | AT405025B (ru) |
AU (1) | AU754824B2 (ru) |
CA (1) | CA2301570A1 (ru) |
EA (1) | EA001640B1 (ru) |
WO (1) | WO1999028740A1 (ru) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE19823814A1 (de) * | 1998-05-27 | 1999-12-02 | Octapharma Ag Ziegelbruecke | Verfahren zur Trennung und/oder Isolierung von Plasmaproteinen mit annularer Chromatographie |
EP1300415B1 (en) | 2001-10-03 | 2007-01-24 | Boehringer Ingelheim International GmbH | Method for reconstituting a biologically active recombinant protein |
CN110441428B (zh) * | 2019-08-19 | 2021-08-24 | 江南大学 | 一种快速分析蛋白质及强极性长氨基酸序列糖肽的方法 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE950281C (de) * | 1949-12-23 | 1956-10-18 | Lkb Produkter Fabriks Aktiebol | Verfahren und Vorrichtung zum chromatographischen Trennen von Gemischen verschiedener Stoffe in Loesungen mittels Sorption |
DE2440848A1 (de) * | 1974-08-26 | 1976-03-18 | W Killer Ag Dr | Verfahren zum fortlaufenden trennen der komponenten eines stoffes und vorrichtung zum durchfuehren des verfahrens |
US4642169A (en) * | 1984-08-01 | 1987-02-10 | University Of Iowa Research Foundation | Continuous rotating electrophoresis column and process of using |
US4740306A (en) * | 1986-03-17 | 1988-04-26 | Temple University - Of The Commonwealth System Of Higher Education | Chromatographic column |
US4683042A (en) * | 1986-04-29 | 1987-07-28 | The United States Of America As Represented By The United States Department Of Energy | Method and apparatus for continuous annular electrochromatography |
US5217608A (en) * | 1988-07-21 | 1993-06-08 | The Research Foundation Of State University Of New York | Multi-column planet centrifuge chromatograph |
US5149436A (en) * | 1988-11-28 | 1992-09-22 | Union Oil Company Of California | Continuous displacement chromatographic method |
DE4231327A1 (de) * | 1992-09-18 | 1994-03-24 | Reuter Karl Dr | Kontinuierliche Chromatographie |
-
1997
- 1997-12-01 AT AT203097A patent/AT405025B/de not_active IP Right Cessation
-
1998
- 1998-11-30 EA EA199901022A patent/EA001640B1/ru not_active IP Right Cessation
- 1998-11-30 CA CA002301570A patent/CA2301570A1/en not_active Abandoned
- 1998-11-30 WO PCT/AT1998/000290 patent/WO1999028740A1/de not_active Application Discontinuation
- 1998-11-30 EP EP98958716A patent/EP1183531A1/de not_active Withdrawn
- 1998-11-30 AU AU14743/99A patent/AU754824B2/en not_active Ceased
Non-Patent Citations (1)
Title |
---|
See references of WO9928740A1 * |
Also Published As
Publication number | Publication date |
---|---|
EA199901022A1 (ru) | 2000-04-24 |
AT405025B (de) | 1999-04-26 |
WO1999028740A1 (de) | 1999-06-10 |
AU754824B2 (en) | 2002-11-28 |
AU1474399A (en) | 1999-06-16 |
EA001640B1 (ru) | 2001-06-25 |
ATA203097A (de) | 1998-09-15 |
CA2301570A1 (en) | 1999-06-10 |
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