WO1999042511A1 - Immobilization of membrane receptor on hplc - Google Patents
Immobilization of membrane receptor on hplc Download PDFInfo
- Publication number
- WO1999042511A1 WO1999042511A1 PCT/US1999/003778 US9903778W WO9942511A1 WO 1999042511 A1 WO1999042511 A1 WO 1999042511A1 US 9903778 W US9903778 W US 9903778W WO 9942511 A1 WO9942511 A1 WO 9942511A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- receptors
- column
- supports
- receptor
- test
- Prior art date
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
- B01D15/26—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
- 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
- B01J20/28033—Membrane, sheet, cloth, pad, lamellar or mat
-
- 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
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to 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
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
- B01J20/3204—Inorganic carriers, supports or substrates
-
- 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
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
-
- 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
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3244—Non-macromolecular compounds
- B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
-
- 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
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3244—Non-macromolecular compounds
- B01J20/3246—Non-macromolecular compounds having a well defined chemical structure
- B01J20/3248—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
- B01J20/3251—Non-macromolecular compounds having a well defined chemical structure the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
-
- 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
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3268—Macromolecular compounds
- B01J20/3272—Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
- B01J20/3274—Proteins, nucleic acids, polysaccharides, antibodies or antigens
-
- 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
-
- 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
- B01J2220/58—Use in a single column
-
- 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
- G01N2030/524—Physical parameters structural properties
- G01N2030/527—Physical parameters structural properties sorbent material in form of a membrane
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
-
- 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
- G01N30/89—Inverse chromatography
Definitions
- This invention relates to immobilization of receptors on a support in a liquid chromatographic system.
- BP-SPs immobilized biopolymer-based LC stationary phases
- This invention provides a immobilized receptors on supports in liquid chromatographic systems. Using the methods of the invention, it is possible to immobilize receptors on supports, then expose those receptors to agents that might attach to the receptors. It is then possible to expose the supports with the receptors, followed by liquid chromatographic studies to determine whether attachment of the agent to the receptor has occurred. It is, of course, also possible to expose the receptors on the supports to substances that might inhibit interaction between the agent that is known to interact with the receptor, then expose the supports with the receptors to the agent to determine whether or not the proposed inhibitor will, in fact, inhibit attachment to the receptor. Hence, using means of the invention, it is possible to test interaction of potential drugs and receptors and also to evaluate possible agents for inhibition of receptor interactions. Description of the Invention:
- membrane receptors play an important role in drug activity and are key targets in combinatorial screens, they have not been included in LC systems. This has been due, in part, to the disruption of the tertiary structure of the receptor produced by covalent immobilization on a solid LC support.
- One solution to this problem is the immobilization of membrane receptors in the phospholipid monolayer of an immobilized artificial membrane (IAM) LC stationary phase.
- IAM immobilized artificial membrane
- the method provides means of evaluating the attachment of agents to receptors comprising the steps of:
- the IAM LC stationary phase is derived from the covalent immobilization of l-myristoyl-2-[ (13-car- boxyl) tridecanoyl] -sn-3-glycerophosphocholine on aminopropyl silica and resembles 1/2 of a cellular membrane.
- Nicotinic acetylcholine receptors are ligand- gated ion channels formed from five homologous subunits oriented like barrel staves center pore. The nAChRs are the primary excitatory neurotransmitter receptors on skeletal muscles and autonomic ganglia in the peripheral nervous system of vertebrates.
- nAChRs In the central nervous system, the nAChRs play important roles in modulating functions of other neurotrans- mitters. Sixteen different subunits of nAChRs have been identified so far. These subunits combine to form a variety of nAChR subtypes.
- the rat ⁇ 3B4 nAChR therefore, was selected for the initial development of a receptor-bearing LC stationary phase.
- the a3/B4 nAChRs prepared from the cell line were solubi- lized using a detergent solution.
- the resulting detergent- receptor solution was mixed with the dry IAM LC support and then dialyzed against Tris-HCl buffer [50 mM, pH 7.4]. Additional buffer was added to the IAM support and the mixture was vortexed, centrifuged and the supernatant decanted.
- the resulting IAM LC support contained approximately 60 g protein per gram IAM support.
- the ability of the immobilized receptors to bind known nAChR ligands was determined using a [ 3 H]epibatidine binding assay protocol designed for cell membrane homogenates.
- the 3/B4 IAM support suspension showed 98% specific binding with 5 r?M active receptor per gram immobilized protein, comparable to 100% specific binding with 8.6 ⁇ K per gram protein found in parallel experiments for the receptor- detergent solution. No specific binding was found to the native IAM LC support.
- the cultured cells in which rat ⁇ 3/B4 subtype of neural nicotinic acetylcholine receptor (nAChR) were expressed by transfected cell line KX 3B4R 2 were harvested in 30 ml of Tris- HC1 buffer (50 mM, pH 7.4) and homogenized for 20 minutes with a Brinkman Polytron homogenizer. The homogenates were centrifuged at 35,000 x g for 10 minutes and the supernatant was discarded. The resulting pellets were suspended in 6 ml of 2% cholate in Tris-HCl buffer (50 mM, pH 7.4) and stirred for two hours in an ice bath.
- Tris- HC1 buffer 50 mM, pH 7.4
- IAM packing materials 200 mg were washed three times by centrifugation with Tris-HCl buffer (50 mM, pH 7.4) .
- the IAM materials were suspended in 4 ml of receptor-detergent solution, stirred for one hour at room temperature and dialyzed against 2 x 1 L buffer for 24 hours at 4°C. When detergents were depleted from the mixture, the receptors were hydrophobically entrapped in the phospholipid monolayer on the surface of IAM materials.
- the receptor-IAM materials were washed with buffer by centrifu- gation at 4000 x g.
- the NR(a4B2)-IAM packing materials were packed in an HR glass column (Pharmacia Biotech) and connected with an HPLC pump. 15-20 ml of [ 3 H]-EB in buffer or in the same buffer containing drugs was applied by a 25 ml sample superloop (Pharmacia Biotech, Uppsala, Sweden) and run at a flow rate of 0.4 ml/min at room temperature. The elution profile was monitored by an on-line radioactivity HPLC detector.
- Example 1 A 0.5 x 1.25 cm LC column with a 0.245 ml bed volume was packed with the (a3/B4) NR-IAM material.
- the resulting column was used in the LC study of drug-NR binding using frontal chromatographic techniques with [ 3 H] -epibatidine (EB) as the marker ligand.
- EB epibatidine
- a series of [ 3 H]-EB concentrations ranging from 60 pM to 600 pM were pumped through the ( ⁇ 3/B4) NR-IAM column2 to obtain elution profiles showing front and plateau regions.
- the concentration of the [ 3 H]-EB in the column eluent was monitored by an LC on-line radioactivity detector.
- the retention volumes of [ 3 H]-EB was increased to 600 pM.
- the binding of the [ 3 H]-EB on the ( ⁇ 3/B4)NR -IAM column was be altered by the addition of competitive NR-ligands to the mobile phase,
- the retention volume of 60 pM [ 3 H]-EB (60 pM) was decreased from 9.5 to 6.6 ml when a 10 nM concentration of the NR-ligand A85380 was added to the mobile phase and fell to
- the relative affinities of ligands for the receptors can be readily classified by determining the concentrations required to decrease the retention volumes of [ 3 H]-EB to a predetermined level. For example, to decrease the retention volumes of 60 pM [ 3 H]-EB from 9.5 ml to 6 ml required mobile phase concentration of 120 pM of ( ⁇ )-EB, 17 mM of A85380, 45 nM of nicotine, 1.3 ⁇ M of carbachol and 21 ⁇ M of atropine, respectively.
- the relative affinities for the NR determined by this method were (+) -EB>A85380>nicotine>carbachol>atropine, which is consistent with results from standard by binding assays.
- Frontal chromatography can be used to calculate dissociation constants, K d , for the marker and displace ligands.
- dissociation constants K d
- the mobile phase concentrations of [ 3 H ⁇ -EB and the competitive ligands were varied and association constant of EB, K EB and the test ligands, K d , as well as the number of the active and available binding sites of immobilized receptors, P, were calculated using equations 1 and 2
- V m a ⁇ - V ) "l ( l+ [ EB ] K EB)( V m i ⁇ [ EB ] K EB) "1+ ( l+ t EB ] K EB- 2 ( V m i ⁇ [ EB ] D EB K drug)- 1
- V - V m i n V m i n [ P ] J EB ) " 1 + ( mi [ Pr 1 [ EB] ( 2 )
- V is the retention volume of EB
- V ma ⁇ is the retention volume of EB at low concentration (60 pM) and in the absence of drugs
- V m1n is the retention volume of EB when the specific interaction is completely suppressed.
- V . was determined by running [ 3 H]-EB in a series of concentration of drugs and plotting 1/ (V ⁇ -V) versus l/[drug] extrapolating to finite [drug] .
- the supports with the receptors may be exposed to drugs or inhibitors, then to drugs followed by chromatographic evaluation of the presence of the drug by chromatographic means to determine whether the drug is present on the support.
- chromatographic means to determine whether proposed inhibitors of receptor/toxin interaction will , in fact, prevent that interaction by exposing the support with receptors bound thereto to proposed inhibitors, then to the toxin or drug followed by chromatographic evaluation of the support to determine whether the toxin or drug has been prevented from binding to the receptor by the inhibitor under consideration.
- any receptor system may be used.
- Other receptors such as GABA receptors have been tested and found to function in a manner similar to those exemplified herein. Examles of other receptors include, steroid receptors such as estrogen based receptors or non-estrogen based receptors that may be turned on by compounds with estrogenic properties would be appropriate for use in the methods of the invention. Androgen receptors could be used to detect possible androgenic activity of substances.
- Supports such as hydrogel beads or hydrophilic verticle support systems may be used in the methods of the invention.
- the receptor binding column can be reused repeatedly
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000532462A JP2002504665A (en) | 1998-02-23 | 1999-02-23 | Immobilization of membrane receptors on HPLC |
EP99908336A EP1064316A4 (en) | 1998-02-23 | 1999-02-23 | Immobilization of membrane receptor on hplc |
CA002322011A CA2322011A1 (en) | 1998-02-23 | 1999-02-23 | Immobilization of membrane receptor on hplc |
AU27795/99A AU765548B2 (en) | 1998-02-23 | 1999-02-23 | Immobilization of membrane receptor on HPLC |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7574598P | 1998-02-23 | 1998-02-23 | |
US60/075,745 | 1998-02-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1999042511A1 true WO1999042511A1 (en) | 1999-08-26 |
Family
ID=22127734
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1999/003778 WO1999042511A1 (en) | 1998-02-23 | 1999-02-23 | Immobilization of membrane receptor on hplc |
Country Status (6)
Country | Link |
---|---|
US (3) | US6139735A (en) |
EP (1) | EP1064316A4 (en) |
JP (1) | JP2002504665A (en) |
AU (1) | AU765548B2 (en) |
CA (1) | CA2322011A1 (en) |
WO (1) | WO1999042511A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1369690A1 (en) * | 2002-06-05 | 2003-12-10 | RETT Corporation | Immobilised G-protein coupled receptors on an artificial lipid membrane of a liquid chromatography stationary phase and use thereof for screening |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000037932A2 (en) * | 1998-12-23 | 2000-06-29 | Admetric Biochem Inc. | High throughput method for measuring physicochemical values |
US6547979B1 (en) | 2000-08-31 | 2003-04-15 | Micron Technology, Inc. | Methods of enhancing selectivity of etching silicon dioxide relative to one or more organic substances; and plasma reaction chambers |
AU2002366956A1 (en) * | 2001-12-19 | 2003-07-09 | Rett Corporation | Novel parallel throughput system |
US20040204862A1 (en) * | 2003-04-11 | 2004-10-14 | Wainer Irving W. | Computer-based model for identification and characterization for non-competitive inhibitors of nicotinic acetylcholine receptors and related ligand-gated ion channel receptors |
FR2867776B1 (en) * | 2004-03-17 | 2006-06-23 | Saint Gobain Vetrotex | GLASS YARNS FOR REINFORCING ORGANIC AND / OR INORGANIC MATERIALS |
US20060226082A1 (en) * | 2005-03-15 | 2006-10-12 | Mcmaster University | Methods for substrate and modulator screening using enzyme-reactor chromatography/tandem mass spectrometry |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4931498A (en) * | 1988-02-25 | 1990-06-05 | Purdue Research Foundation | Immobilized artificial membranes |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4693985A (en) * | 1984-08-21 | 1987-09-15 | Pall Corporation | Methods of concentrating ligands and active membranes used therefor |
EP0235526A3 (en) * | 1986-01-29 | 1988-03-23 | Leipziger Arzneimittelwerk GmbH | Activated polymer solids and process for their manufacture |
US4927879A (en) * | 1988-02-25 | 1990-05-22 | Purdue Research Foundation | Method for solid phase membrane mimetics |
US5340474A (en) * | 1988-03-24 | 1994-08-23 | Terrapin Technologies, Inc. | Panels of analyte-binding ligands |
US5045190A (en) * | 1988-11-08 | 1991-09-03 | Carbonell Ruben G | Chromatography apparatus |
US5149425A (en) * | 1988-11-09 | 1992-09-22 | Chembiomed, Ltd. | Affinity supports for hemoperfusion |
US5240601A (en) * | 1988-11-09 | 1993-08-31 | Chembiomed, Ltd. | Affinity supports for hemoperfusion |
US4957620A (en) * | 1988-11-15 | 1990-09-18 | Hoechst Celanese Corporation | Liquid chromatography using microporous hollow fibers |
US5160627A (en) * | 1990-10-17 | 1992-11-03 | Hoechst Celanese Corporation | Process for making microporous membranes having gel-filled pores, and separations methods using such membranes |
US5240856A (en) * | 1991-10-23 | 1993-08-31 | Cellpro Incorporated | Apparatus for cell separation |
US5529686A (en) * | 1994-07-15 | 1996-06-25 | Minnesota Mining And Manufacturing Company | Composite membranes for solid phase extractions and reactions |
JPH11513568A (en) * | 1995-10-17 | 1999-11-24 | ジェネンコア インターナショナル インコーポレーテッド | Enzyme sequence and method for producing the same |
-
1999
- 1999-02-23 US US09/255,881 patent/US6139735A/en not_active Expired - Fee Related
- 1999-02-23 WO PCT/US1999/003778 patent/WO1999042511A1/en not_active Application Discontinuation
- 1999-02-23 CA CA002322011A patent/CA2322011A1/en not_active Abandoned
- 1999-02-23 AU AU27795/99A patent/AU765548B2/en not_active Ceased
- 1999-02-23 JP JP2000532462A patent/JP2002504665A/en active Pending
- 1999-02-23 EP EP99908336A patent/EP1064316A4/en not_active Withdrawn
-
2000
- 2000-07-19 US US09/619,505 patent/US6387268B1/en not_active Expired - Fee Related
-
2002
- 2002-05-13 US US10/142,785 patent/US20020125194A1/en not_active Abandoned
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4931498A (en) * | 1988-02-25 | 1990-06-05 | Purdue Research Foundation | Immobilized artificial membranes |
Non-Patent Citations (4)
Title |
---|
BARBATO, F. LA ROTONDA, M. QUAGLIA, F.: "Chromatographic indices determined on an immobilized artificial membrane (IAM) column as descriptors of lipophilic and polar interactions of 4-phenyldihydropyridine calcium-channel blockers with biomembranes", EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY., EDITIONS SCIENTIFIQUE ELSEVIER, PARIS., FR, vol. 31, no. 4, 1 January 1996 (1996-01-01), FR, pages 311 - 318, XP004040091, ISSN: 0223-5234, DOI: 10.1016/0223-5234(96)80368-0 * |
IM W. B., BLAKEMAN D. P., DAVIS J. P.: "IMMOBILIZED GABAA RECEPTORS AND THEIR LIGAND BINDING CHARACTERISTICS.", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ACADEMIC PRESS INC. ORLANDO, FL, US, vol. 163., no. 01., 30 August 1989 (1989-08-30), US, pages 611 - 617., XP002920894, ISSN: 0006-291X, DOI: 10.1016/0006-291X(89)92181-5 * |
KALISZAN, R. NASAL, A. TUROWSKI, M.: "Quantitative structure-retention relationships in the examination of the topography of the binding site of antihistamine drugs on @a"1-acid glycoprotein", JOURNAL OF CHROMATOGRAPHY, ELSEVIER SCIENCE PUBLISHERS B.V., NL, vol. 722, no. 1, 26 January 1996 (1996-01-26), NL, pages 25 - 32, XP004038260, ISSN: 0021-9673, DOI: 10.1016/0021-9673(95)00523-4 * |
See also references of EP1064316A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1369690A1 (en) * | 2002-06-05 | 2003-12-10 | RETT Corporation | Immobilised G-protein coupled receptors on an artificial lipid membrane of a liquid chromatography stationary phase and use thereof for screening |
Also Published As
Publication number | Publication date |
---|---|
JP2002504665A (en) | 2002-02-12 |
US6139735A (en) | 2000-10-31 |
EP1064316A1 (en) | 2001-01-03 |
CA2322011A1 (en) | 1999-08-26 |
US6387268B1 (en) | 2002-05-14 |
EP1064316A4 (en) | 2002-07-31 |
US20020125194A1 (en) | 2002-09-12 |
AU2779599A (en) | 1999-09-06 |
AU765548B2 (en) | 2003-09-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Pap et al. | Effect of solvents on the selectivity of terbutylazine imprinted polymer sorbents used in solid-phase extraction | |
Takeuchi et al. | Separation and sensing based on molecular recognition using molecularly imprinted polymers | |
Zhu et al. | Selective solid-phase extraction using molecularly imprinted polymer for the analysis of polar organophosphorus pesticides in water and soil samples | |
Schomburg et al. | Immobilization of stationary liquids on silica particles by γ-radiation | |
Podgornik et al. | High-performance membrane chromatography of small molecules | |
Gunasena et al. | Organic monoliths for hydrophilic interaction electrochromatography/chromatography and immunoaffinity chromatography | |
EP0290406B1 (en) | Low affinity adsorbent for affinity chromatography | |
Yu et al. | Macroporous poly (glycidyl methacrylate–triallyl isocyanurate–divinylbenzene) matrix as an anion-exchange resin for protein adsorption | |
US6139735A (en) | Immobilization of membrane receptor on HPLC | |
EP2254696B1 (en) | Improved chromatography resin, and methods and devices related thereto | |
Hermansson | Direct liquid chromatographic resolution of racemic drugs by means of α1-acid glycoprotein as the chiral complexing agent in the mobile phase | |
Kasche et al. | Resolution in high-performance liquid affinity chromatography: Dependence on eluite diffusion into the stationary phase | |
Davidson et al. | Molecular imprinting of biologically active steroidal systems | |
Cichna-Markl | Selective sample preparation with bioaffinity columns prepared by the sol–gel method | |
Nash et al. | Modification of polystyrenic matrices for the purification of proteins: II. Effect of the degree of glutaraldehyde–poly (vinyl alcohol) crosslinking on various dye ligand chromatography systems | |
EP2830755B1 (en) | Method for reverse phase chromatography of active pharmaceutical ingredients | |
Loukili et al. | Study of tryptophan enantiomer binding to a teicoplanin-based stationary phase using the perturbation technique: investigation of the role of sodium perchlorate in solute retention and enantioselectivity | |
Gasparrini et al. | New chiral and restricted-access materials containing glycopeptides as selectors for the high-performance liquid chromatographic determination of chiral drugs in biological matrices | |
Messina et al. | Direct resolution of optically active isomers on chiral packings containing ergoline skeletons. 5. Enantioseparation of amino acid derivatives | |
Maier et al. | Stereoselective chromatographic methods for drug analysis | |
Claessens | Characterization of stationary phases for reversed-phase liquid chromatography: column testing, classification and chemical stability | |
Gasparrini et al. | Synthesis and characterization of novel internal surface reversed-phase silica supports for high-performance liquid chromatography | |
Akapo et al. | Retention characteristics and selected applications of cyclic siloxane-based octadecylsilyl bonded phases in reversed-phase high-performance liquid chromatography | |
Gomez et al. | Surface modification on poly (EGDMA-co-HEMA) synthetic matrices to be used as specific adsorbents | |
Hollis et al. | Fast affinity chromatography using small particle silica-based packing materials |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AU CA GB JP |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
ENP | Entry into the national phase |
Kind code of ref document: A Ref document number: 2322011 Country of ref document: CA Ref document number: 2000 532462 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1999908336 Country of ref document: EP Ref document number: 27795/99 Country of ref document: AU |
|
WWP | Wipo information: published in national office |
Ref document number: 1999908336 Country of ref document: EP |
|
WWG | Wipo information: grant in national office |
Ref document number: 27795/99 Country of ref document: AU |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1999908336 Country of ref document: EP |