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1. WO2004024950 - METHOD FOR THE SELECTION OF NUCLEIC ACID LIGANDS

Publication Number WO/2004/024950
Publication Date 25.03.2004
International Application No. PCT/EP2003/010053
International Filing Date 10.09.2003
IPC
B01D 15/08 2006.01
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
15Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
08Selective adsorption, e.g. chromatography
B01D 15/38 2006.01
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
15Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
08Selective adsorption, e.g. chromatography
26characterised by the separation mechanism
38involving specific interaction not covered by one or more of groups B01D15/30-B01D15/36120
B01D 61/14 2006.01
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
61Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
14Ultrafiltration; Microfiltration
B01D 61/16 2006.01
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
61Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
14Ultrafiltration; Microfiltration
16Feed pretreatment
B01L 3/00 2006.01
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
3Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
C12N 15/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
CPC
B01D 15/3804
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
15Separating processes involving the treatment of liquids with solid sorbents
08Selective adsorption, e.g. chromatography
26characterised by the separation mechanism
38involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
3804Affinity chromatography
B01D 2311/04
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
2311Details relating to membrane separation process operations and control
04Specific process operations in the feed stream; Feed pretreatment
B01D 61/145
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
61Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis, ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
14Ultrafiltration; Microfiltration
145Ultrafiltration
B01D 61/16
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
DSEPARATION
61Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis, ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
14Ultrafiltration; Microfiltration
16Feed pretreatment
B01L 3/50255
BPERFORMING OPERATIONS; TRANSPORTING
01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
3Containers or dishes for laboratory use, e.g. laboratory glassware
50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
502with fluid transport, e.g. in multi-compartment structures
5025for parallel transport of multiple samples
50255Multi-well filtration
C12N 15/1048
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
1034Isolating an individual clone by screening libraries
1048SELEX
Applicants
  • NOXXON PHARMA AG [DE]/[DE] (AllExceptUS)
  • EULBERG, Dirk [DE]/[DE] (UsOnly)
  • KLUSSMANN, Sven [DE]/[DE] (UsOnly)
Inventors
  • EULBERG, Dirk
  • KLUSSMANN, Sven
Agents
  • BOHMANN, Armin, K.
Priority Data
102 41 938.810.09.2002DE
Publication Language German (DE)
Filing Language German (DE)
Designated States
Title
(DE) VERFAHREN ZUR SELEKTION VON NUKLEINSÄURELIGANDEN
(EN) METHOD FOR THE SELECTION OF NUCLEIC ACID LIGANDS
(FR) PROCEDE DE SELECTION D'ACIDES NUCLEIQUES LIGANDS
Abstract
(DE)
Die vorliegende Erfindung betrifft ein Verfahren zur Selektion eines oder mehrerer Nukleinsäureliganden aus einer Mischung von Kandidaten-Nukleinsäuren, wobei die Nukleinsäureliganden an ein Zielmolekül binden können, umfassend die Schritte a) Kontaktieren der Mischung mit dem immobilisierten Zielmolekül, b) Abtrennen der mit einer höheren Affinität an das Zielmolekül bindende(n) Nukleinsäure(n) vom Rest der Kandidatenmischung, wobei die mit einer höheren Affinität an das Zielmolekül bindenden Nukleinsäure(n) in einem Komplex mit dem Zielmolekül vorliegen, c) Elution der mit einer höheren Affinität an das Zielmolekül bindenden Nukleinsäure(n) von dem Zielmolekül; und d) optional Amplifizieren der mit höherer Affinität bindenden Nukleinsäure(n), wodurch ein angereichertes Nukleinsäureprodukt entsteht, wobei das Zielmolekül an einer Oberfläche immobilisiert vorliegt und das Verfahren einen Ultrafiltrationsschritt umfasst.
(EN)
The invention relates to a method for the selection of one or more nucleic acid ligands from a mixture of candidate - nucleic acids. The nucleic acid ligands can bind to a target molecule. Said inventive method comprises the following steps a) placing the mixture in contact with the immobilised target molecule, b) separating the nucleic acid(s) binding to the target molecule with a higher affinity from the remainder of the candidate mixture, whereby the nucleic acid(s) binding to the target molecule with a higher affinity are provided in a complex with the target molecule, c) eluting the nucleic acid(s) binding to the target molecule with a higher affinity from the target molecule and d) optionally amplifying the binding nucleic acid(s) with a higher affinity, whereby an enriched nucleic acid product is produced. The target molecule is immobilised on the surface and the method also comprises an ultra filtration step.
(FR)
La présente invention concerne un procédé de sélection d'au moins un acide nucléique ligand à partir d'un mélange d'acides nucléiques candidats, les acides nucléiques ligands pouvant se fixer à une molécule cible. Ce procédé comprend les étapes suivantes: a) mise en contact du mélange avec la molécule cible immobilisée; b) séparation de l'acide ou des acides nucléiques se liant avec une affinité élevée à la molécule cible du reste du mélange de candidat, l'acide ou les acides nucléiques se fixant avec une affinité élevée à la molécule cible formant un complexe avec celle-ci; c) élution de la molécule cible de l'acide ou des acides nucléiques se fixant à celle-ci avec une affinité élevée; et d) amplification éventuelle de l'acide ou des acides nucléiques se fixant avec une affinité élevée, cette amplification permettant d'obtenir un acide ou des acides nucléiques enrichis. La molécule cible utilisée est immobilisée sur une surface et le procédé comprend une étape d'ultrafiltration.
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