Method for conducting receptor-ligand association reaction and reactor used therefor

• US 20030148402A1
• Filed: 01/27/2003
• Published: 08/07/2003
• Est. Priority Date: 02/01/2002
• Status: Abandoned Application

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Amano, Yoshikazu, Tsuzuki, Hirohiko
• Application Number:
  US10/351,379
• Publication Number:
  US 20030148402A1
• Time in Patent Office: Days
• US Class Current: 435/7.9
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00578 electrophoric
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0605 : check valves
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: C12Q 1/6832 : Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2565/137 : Chromatographic separation
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing

Abstract

A method for conducting a receptor-ligand association reaction includes the steps of forcibly feeding a reaction solution containing a ligand labeled with a labeling substance from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing receptors toward the other side thereof so as to cut through the absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively associating the ligand contained in the reaction solution with the receptors contained in the absorptive regions. According to this method, it is possible to efficiently associate a ligand with receptors fixed in the absorptive regions of the biochemical analysis unit and produce biochemical analysis data having an excellent quantitative characteristic with good repeatability.

Claims

1. A method for conducting a receptor-ligand association reaction comprising the steps of forcibly feeding a reaction solution containing a ligand or receptor labeled with a labeling substance from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing receptors or ligands toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively associating the ligand or receptor contained in the reaction solution with the receptors or the ligands contained in the plurality of absorptive regions of the biochemical analysis unit.

2. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the reaction solution contains a ligand or receptor labeled with a labeling substance selected from a group consisting of a radioactive labeling substance, a fluorescent substance and a labeling substance which generates chemiluminescence emission when it contacts a chemiluminescent substrate.

3. A method for conducting a receptor-ligand association reaction in accordance with claim 1 which comprises steps of forcibly feeding a reaction solution containing a substance derived from a living organism and labeled with a labeling substance from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing specific binding substances toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively associating the substance derived from a living organism with the specific binding substances contained in the plurality of absorptive regions of the biochemical analysis unit.

4. A method for conducting a receptor-ligand association reaction in accordance with claim 3 which comprises steps of forcibly feeding a reaction solution containing a substance derived from a living organism and labeled with a labeling substance from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing specific binding substances whose structure or characteristics are known toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively hybridizing the substance derived from a living organism, labeled with the labeling substance and contained in the reaction solution with the specific binding substances contained in the plurality of absorptive regions of the biochemical analysis unit.

5. A method for conducting a receptor-ligand association reaction in accordance with claim 4 which comprises steps of forcibly feeding a reaction solution containing a substance derived from a living organism and labeled with a hapten from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing specific binding substances whose structure or characteristics are known toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively hybridizing the substance derived from a living organism, labeled with the hapten and contained in the reaction solution with the specific binding substances contained in the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding a reaction solution containing an antibody to the hapten labeled with a labeling enzyme from the one side of the biochemical analysis unit toward the other side thereof, and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby binding by an antigen-antibody reaction the antibody labeled with the labeling enzyme with the hapten contained in the plurality of absorptive regions of the biochemical analysis unit.

6. A method for conducting a receptor-ligand association reaction in accordance with claim 5 which comprises steps of forcibly feeding a reaction solution containing a substance derived from a living organism and labeled with a hapten from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing specific binding substances whose structure or characteristics are known toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively hybridizing the substance derived from a living organism, labeled with the hapten and contained in the reaction solution with the specific binding substances contained in the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding a reaction solution containing an antibody to the hapten labeled with an enzyme which generates chemiluminescence emission when it contacts a chemiluminescent substrate from the one side of the biochemical analysis unit toward the other side thereof, and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby binding by an antigen-antibody reaction the antibody labeled with the enzyme with the hapten contained in the plurality of absorptive regions of the biochemical analysis unit.

7. A method for conducting a receptor-ligand association reaction in accordance with claim 5 which comprises steps of forcibly feeding a reaction solution containing a substance derived from a living organism and labeled with a hapten from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing specific binding substances whose structure or characteristics are known toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby selectively hybridizing the substance derived from a living organism, labeled with the hapten and contained in the reaction solution with the specific binding substances contained in the plurality of absorptive regions of the biochemical analysis unit, forcibly feeding a reaction solution containing an antibody to the hapten labeled with an enzyme which generates a fluorescent substance when it contacts a fluorescent substrate from the one side of the biochemical analysis unit toward the other side thereof, and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby binding by an antigen-antibody reaction the antibody labeled with the enzyme with the hapten contained in the plurality of absorptive regions of the biochemical analysis unit.

8. A method for conducting a receptor-ligand association reaction in accordance with claim 1 which comprises steps of forcibly feeding a reaction solution containing an antibody bound with a labeling enzyme from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing an antigen labeling a substance derived from a living organism selectively hybridized with specific binding substances toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby associating the antibody bound with the labeling enzyme with the antigen contained in the plurality of absorptive regions of the biochemical analysis unit by an antigen-antibody reaction.

9. A method for conducting a receptor-ligand association reaction in accordance with claim 8 which comprises steps of forcibly feeding a reaction solution containing an antibody or an antigen labeled with a labeling substance from one side of a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing an antigen or an antibody toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via at least one check valve, thereby binding by an antigen-antibody reaction the antibody or the antigen labeled with a labeling substance and contained in the reaction solution with the antigen or the antibody contained in the plurality of absorptive regions of the biochemical analysis unit.

10. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the reaction solution is forcibly fed using at least one syringe provided with a piston from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by the syringe from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

11. A method for conducting a receptor-ligand association reaction in accordance with claim 4 wherein the reaction solution is forcibly fed using at least one syringe provided with a piston from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by the syringe from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

12. A method for conducting a receptor-ligand association reaction in accordance with claim 6 wherein the reaction solution is forcibly fed using at least one syringe provided with a piston from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by the syringe from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

13. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the reaction solution is forcibly fed by deforming two diaphragms facing each other from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by deforming the diaphragms from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

14. A method for conducting a receptor-ligand association reaction in accordance with claim 4 wherein the reaction solution is forcibly fed by deforming two diaphragms facing each other from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by deforming the diaphragms from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

15. A method for conducting a receptor-ligand association reaction in accordance with claim 6 wherein the reaction solution is forcibly fed by deforming two diaphragms facing each other from the one side of a biochemical analysis unit toward the other side thereof so as to cut through the plurality of absorptive regions of the biochemical analysis unit and the reaction solution is forcibly fed by deforming the diaphragms from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

16. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the biochemical analysis unit includes a substrate formed with a plurality of holes spaced apart from each other and the plurality of absorptive regions are formed by charging an absorptive material in the plurality of holes formed in the substrate and causing the absorptive material charged in the plurality of holes to contain receptors or ligands.

17. A method for conducting a receptor-ligand association reaction in accordance with claim 16 wherein the plurality of absorptive regions are formed by pressing an absorptive membrane containing an absorptive material into the plurality of through-holes formed in the substrate and causing the absorptive material charged in the plurality of through-holes to contain receptors or ligands.

18. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the biochemical analysis unit is formed with 10 or more absorptive regions.

19. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein each of the plurality of absorptive regions of the biochemical analysis unit has a size of less than 5 mm².

20. A method for conducting a receptor-ligand association reaction in accordance with claim 1 wherein the plurality of absorptive regions are formed in the biochemical analysis unit at a density of 10 or more per cm².

21. A method for conducting a receptor-ligand association reaction in accordance with claim 16 wherein the substrate of the biochemical analysis unit preferably has a property of attenuating radiation energy.

22. A method for conducting a receptor-ligand association reaction in accordance with claim 21 wherein the substrate of the biochemical analysis unit has a property of reducing the energy of radiation to ⅕

or less when the radiation travels in the substrate by a distance equal to that between neighboring absorptive regions.

23. A method for conducting a receptor-ligand association reaction in accordance with claims 16 wherein the substrate of the biochemical analysis unit preferably has a property of attenuating light energy.

24. A method for conducting a receptor-ligand association reaction in accordance with claim 23 wherein the substrate of the biochemical analysis unit has a property of reducing the energy of light to ⅕

or less when the light travels in the substrate by a distance equal to that between neighboring absorptive regions.

25. A reactor for conducting a receptor-ligand association reaction comprising a reaction vessel provided with a biochemical analysis unit holding section formed with at least one check valve for holding a biochemical analysis unit formed with a plurality of absorptive regions spaced apart from each other and containing receptors or ligands and adapted for accommodating a reaction solution containing a ligand or receptor, and a solution feeding means for forcibly feeding the reaction solution accommodated in the reaction vessel from one side of the biochemical analysis unit held by the biochemical analysis unit holding section toward the other side of the biochemical analysis unit so as to cut through the plurality of absorptive regions of the biochemical analysis unit and forcibly feeding the reaction solution from the other side of the biochemical analysis unit toward the one side thereof via the at least one check valve.

26. A reactor for conducting a receptor-ligand association reaction in accordance with claim 25 wherein the solution feeding means is constituted as at least one syringe provided with a piston.

27. A reactor for conducting a receptor-ligand association reaction in accordance with claim 25 wherein the solution feeding means is constituted as two deferrable diaphragms facing each other.