Assay method using magnetic silica particles and reagent for said assay method

1. A method of assaying an analyte in a sample in which magnetic silica particles are used,

wherein the magnetic silica particles comprise silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and an analyte-binding substance that is a substance capable of specifically binding to the analyte and is immobilized on a surface of the silica particles, and

wherein the method comprises:

bringing a sample containing the analyte, the magnetic silica particles, and a labeled analyte-binding substance that is an analyte-binding substance labeled by a label into contact with each other to form a labeled complex that is a complex of the analyte-binding substance, the analyte, and the labeled analyte-binding substance on the magnetic silica particles;

B/F-separating the magnetic silica particles carrying the labeled complex;

measuring the amount of the label in the labeled complex; and

assaying the analyte in the sample based on the resulting amount of the label.

2. A method of assaying an analyte in a sample in which magnetic silica particles are used,

wherein the magnetic silica particles comprise silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and the analyte or an analog of the analyte immobilized on a surface of the silica particles, and

wherein the method comprises:

bringing a sample containing the analyte, a labeled analyte-binding substance that is a substance labeled by a label and is capable of specifically binding to the analyte, and the magnetic silica particles into contact with each other to form a labeled complex that is a complex of the analyte or the analog of the analyte and the labeled analyte-binding substance on the magnetic silica particles;

B/F-separating the magnetic silica particles carrying the labeled complex;

measuring the amount of the label in the labeled complex; and

assaying the analyte in the sample based on the resulting amount of the label.

3. A method of assaying an analyte in a sample in which magnetic silica particles are used,

wherein the magnetic silica particles comprise silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and an analyte-binding substance that is a substance capable of specifically binding to the analyte immobilized on a surface of the silica particles, and

wherein the method comprises:

bringing a sample containing the analyte, a labeled analyte or analog thereof that is an analyte or analog of the analyte labeled by a label, and the magnetic silica particles into contact with each other to form a labeled complex that is a complex of the analyte-binding substance and the labeled analyte or analog on the magnetic silica particles;

B/F-separating the magnetic silica particles carrying the labeled complex;

measuring the amount of the label in the labeled complex; and

assaying the analyte in the sample based on the resulting amount of the label.

4. The assay method according to claim 1 ,

wherein the B/F separation is performed utilizing the magnetism of the magnetic silica particles.

5. The assay method according to claim 1 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

6. The assay method according to claim 1 ,

wherein the superparamagnetic metal oxide is an iron oxide.

7. The assay method according to claim 6 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

8. The assay method according to claim 1 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the substance capable of specifically binding to the analyte on the surface of the magnetic silica particles.

9. The assay method according to claim 8 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

10. The assay method according to claim 1 ,

wherein the substance capable of specifically binding to the analyte is an antibody against the analyte or an analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.

11. A reagent for the assay method according to claim 1 , comprising

magnetic silica particles comprising:

silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and

a substance capable of specifically binding to an analyte immobilized on a surface of the silica particles.

12. The reagent according to claim 11 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

13. The reagent according to claim 11 ,

wherein the superparamagnetic metal oxide is an iron oxide.

14. The reagent according to claim 13 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

15. The reagent according to claim 11 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the substance capable of specifically binding to the analyte on the surface of the magnetic silica particles.

16. The reagent according to claim 15 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

17. The reagent according to claim 11 ,

wherein the substance capable of specifically binding to the analyte is an antibody against the analyte or an analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.

18. The assay method according to claim 2 ,

wherein the B/F separation is performed utilizing the magnetism of the magnetic silica particles.

19. The assay method according to claim 2 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

20. The assay method according to claim 2 ,

wherein the superparamagnetic metal oxide is an iron oxide.

21. The assay method according to claim 20 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

22. The assay method according to claim 2 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the analyte or the analog of the analyte on the surface of the magnetic silica particles.

23. The assay method according to claim 22 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

24. The assay method according to claim 2 ,

wherein the labeled analyte-binding substance is an antibody against the analyte or the analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.

25. A reagent for the assay method according to claim 2 , comprising

magnetic silica particles comprising:

silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and

an analyte or an analog of the analyte immobilized on a surface of the silica particles.

26. The reagent according to claim 25 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

27. The reagent according to claim 25 ,

wherein the superparamagnetic metal oxide is an iron oxide.

28. The reagent according to claim 27 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

29. The reagent according to claim 25 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the analyte or the analog of the analyte on the surface of the magnetic silica particles.

30. The reagent according to claim 29 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

31. The reagent according to claim 25 ,

comprising a labeled analyte-binding substance that is a substance labeled by a label and is capable of specifically binding to the analyte, wherein the labeled analyte-binding substance is an antibody against the analyte or the analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.

32. The assay method according to claim 3 ,

wherein the B/F separation is performed utilizing the magnetism of the magnetic silica particles.

33. The assay method according to claim 3 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

34. The assay method according to claim 3 ,

wherein the superparamagnetic metal oxide is an iron oxide.

35. The assay method according to claim 34 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

36. The assay method according to claim 3 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the substance capable of specifically binding to the analyte on the surface of the magnetic silica particles.

37. The assay method according to claim 36 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

38. The assay method according to claim 3 ,

wherein the substance capable of specifically binding to the analyte is an antibody against the analyte or an analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.

39. A reagent for the assay method according to claim 3 , comprising

magnetic silica particles comprising:

silica particles containing 60 to 95% by weight of a superparamagnetic metal oxide that has an average particle size of 1 to 15 nm; and

a substance capable of specifically binding to an analyte immobilized on a surface of the silica particles.

40. The reagent according to claim 39 ,

wherein the magnetic silica particles have an average particle size of 1 to 5 μm.

41. The reagent according to claim 39 ,

wherein the superparamagnetic metal oxide is an iron oxide.

42. The reagent according to claim 41 ,

wherein the iron oxide is at least one selected from the group consisting of magnetite, γ-hematite, an intermediate iron oxide between magnetite and α-hematite, and an intermediate iron oxide between γ-hematite and α-hematite.

43. The reagent according to claim 39 ,

wherein at least one organic compound selected from the group consisting of glutaraldehyde, albumin, carbodiimide, streptavidin, biotin, and functional group-containing alkylalkoxysilanes is bound to the surface of the magnetic silica particles before the immobilization of the substance capable of specifically binding to the analyte on the surface of the magnetic silica particles.

44. The reagent according to claim 43 ,

wherein the functional group of the alkylalkoxysilanes is at least one selected from the group consisting of an amino group, a carboxyl group, a hydroxy group, a mercapto group, and a glycidyloxy group.

45. The reagent according to claim 39 ,

wherein the substance capable of specifically binding to the analyte is an antibody against the analyte or an analog of the analyte, an antigen capable of binding to the analyte or the analog of the analyte, or a protein capable of binding to the analyte or the analog of the analyte.