Device for the capture of target molecules, and capturing process using the device

• US 5,723,344 A
• Filed: 06/24/1994
• Issued: 03/03/1998
• Est. Priority Date: 06/25/1993
• Status: Expired due to Fees

• Current Assignee: Bio Merieux
• Original Assignee: Bio Merieux
• Inventors: Mabilat, Claude, Mandrand, Bernard, Pichot, Christian, Charles, Marie-Helene, Cros, Philippe, Erout, Marie-Noelle
• Primary Examiner(s): Hutzell, Paula K.
• Assistant Examiner(s): GRUN, JAMES LESLIE
• Application Number:
  US08/264,996
• Time in Patent Office: 1,348 Days
• Field of Search: 435/6, 435/7.5, 435/7.1, 435/7.92, 435/7.94, 435/975, 435/287.2, 435/287.9, 436/518, 436/524, 436/528, 436/531, 436/532, 436/534, 436/807, 525/54.1, 422/57, 422/60
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

Device for capturing a target molecule for the purpose of detecting it and/or assaying it, including a solid support on which is immobilized a ligand, the ligand being provided in the form of a conjugate resulting from the covalent coupling of a polymer with a plurality of molecules of the ligand. The polymer is an N-vinylpyrrolidone copolymer, and the conjugate is immobilized on the solid support by adsorption.

When the ligand is capable of forming a complex with the target, the device is specific for a given target. When the device comprises, in addition, a bifunctional reagent capable of forming a complex, on the one hand, with the ligand and, on the other hand, with the target, the support on which the ligand is immobilized constitutes a universal capturing system.

Claims

1. Device for capturing a target molecule for the purpose of detecting it and/or assaying it, comprising a water-soluble conjugate immobilized on a solid support by adsorption, said conjugate resulting from a covalent coupling of a polymer with a plurality of molecules of a ligand, wherein said polymer is an N-vinylpyrrolidone copolymer.

2. Device according to claim 1, wherein the copolymer results from copolymerization of an N-vinyl pyrrolidone monomer and a second monomer that establishes a covalent coupling between the ligand and the copolymer.

3. Device according to claim 2, wherein the second monomer contains a reactive functional group or an activable functional group that forms a covalent bond between the copolymer and the ligand, said reactive functional group being selected from the group consisting of aldehyde, epoxy, haloalkyl, primary amine, thiol, maleimide and ester functional groups, and said activable functional group being selected from the group consisting of carboxyl and hydroxyl functional groups.

4. Device according to claim 3, wherein said reactive functional group is a N-hydroxysuccinimide ester functional group.

5. Device according to claim 2, wherein the second monomer is N-acryloxysuccinimide.

6. Device according to claim 1, wherein said copolymer contains from 25 to 70%, in units, of N-vinylpyrrolidone units.

7. Device according to claim 1, wherein said ligand binds with an anti-ligand to form a ligand/anti-ligand capture complex.

8. Device according to claim 7, wherein said anti-ligand is the target molecule.

9. Device according to claim 8, wherein said complex is selected from the group consisting of antigen/antibody, antibody/hapten, hormone/receptor, and chelator/chelated molecule, polynucleotide/polynucleotide and polynucleotide/nucleic acid.

10. Device according to claim 7, wherein said ligand is a polynucleotide containing from 5 to 100 nucleotides.

11. Device according to claim 1, wherein the solid support is selected from the group consisting of a microtitre plate, a sheet, a cone, a tube, a well and a bead.

12. A process for capturing a target molecule, comprising:

bringing a device according to claim 8 into contact with the target molecule in solution in an aqueous medium under conditions permitting the formation of a complex between the ligand and the target; and

allowing said complex to form.

13. Kit for capturing a target molecule for the purpose of detecting and/or assaying it, comprising:

a container containing (1) a water-soluble conjugate immobilized on a solid support by adsorption, said conjugate resulting from a covalent coupling of a polymer with a plurality of molecules of a ligand, wherein said polymer is an N-vinyl pyrrolidone copolymer, and (2) a bifunctional reagent comprising an anti-ligand group bonded with a partner group, wherein said anti-ligand group binds with the ligand to form a complex and said partner group binds with the target to form a partner/target complex.

14. Kit according to claim 13, wherein the anti-ligand group binds with the ligand to form a complex selected from the group consisting of antigen/antibody, antibody/hapten, hormone/receptor, polynucleotide/polynucleotide hybrid, polynucleotide/nucleic acid hybrid, chelator/chelated molecule, biotin/streptavidin and lectin/sugar complexes.

15. A process for capturing a target molecule, comprising:

providing a device comprising;

(1) a water-soluble conjugate immobilized on a solid support by adsorption, said conjugate resulting from a covalent coupling of an N-vinylpyrrolidone copolymer with a plurality of molecules of a ligand, and (2) a bifunctional reagent comprising an anti-ligand group bonded with a partner group, wherein said anti-ligand group is attached to a ligand of the water-soluble conjugate to form a complex;

bringing the device into contact with the target molecule in solution in an aqueous medium under conditions permitting the formation of a complex between the partner group and the target; and

allowing said complex between the partner and the target to form.