Novel material for immunological assay of biochemical components and a process for the determination of said components

• US 4,342,739 A
• Filed: 01/08/1980
• Issued: 08/03/1982
• Est. Priority Date: 01/09/1979
• Status: Expired due to Term

• Current Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Original Assignee: Fuji Photo Film Co Limited (Fujifilm Holdings Corporation)
• Inventors: Kakimi, Fujio, Hiratsuka, Nobuo, Matsukawa, Hiroharu
• Primary Examiner(s): Nucker, Christine M.
• Application Number:
  US06/110,318
• Time in Patent Office: 938 Days
• Field of Search: 424/1, 424/12, 424/32, 23/230 B, 252/316
• US Class Current: 435/7.9
• CPC Class Codes: G01N 2446/20 the magnetic material being present in the particle core
• CPC Class Codes: G01N 2446/30 the magnetic material being dispersed in the polymer composition before their conversion into particulate form
• CPC Class Codes: G01N 2446/90 characterised by small molecule linker used to couple immunoreagents to magnetic particles
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/5434 using magnetic particle immunoreagent carriers which constitute new materials per se
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: Y10S 436/823 Immunogenic carrier or carrier per se
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 428/2984 Microcapsule with fluid core [includes liposome]
• CPC Class Codes: Y10T 428/2985 Solid-walled microcapsule from synthetic polymer

Abstract

Antigen- or antibody-bound microcapsules are employed for immunological assay of biochemical components having immunological activity. The microcapsule system provides high detection sensitivity, high accuracy and excellent reproducibility in immunological assay, without causing non-specific reaction. The microcapsules can be labelled with isotopes, enzymes, etc., for use in quantitative assays of trace amounts of biochemical components.

Claims

1. A microcapsule comprising an oily liquid core material and a wall material having bound thereto an antigen or antibody to the surface thereof, wherein a marking substance is incorporated into said core material.

2. The microcapsule of claim 1 wherein the specific gravity thereof is between about 0.8 and about 1.20.

3. The microcapsule of claim 2 wherein said specific gravity is between 1.05 and 1.20.

4. The microcapsule of claim 3 wherein said specific gravity is between 1.07 and 1.16.

5. The microcapsule of claim 1 wherein said marking substance is selected from the group consisting of an isotope, an enzyme, a fluorescent substance, a UV absorbent and a dye.

6. The microcapsule of claim 5 wherein said isotope is selected from the group consisting of ¹²⁵ I, ¹³¹ I, ³ H and ¹⁴ C.

7. The microcapsule of claim 5 wherein said enzyme is selected from the group consisting of alkali phosphatase, β

-galactosidase, acetylcholine esterase, glucoamylase, maleic acid dehydrogenase glucose-6-phosphoric acid dehydrogenase, peroxidase and glutaroxidase.

8. The microcapsule of claim 5 wherein said fluorescent substance is selected from the group consisting of stilbene and derivatives thereof, coumarine derivatives, pyrazoline derivatives,

9. The microcapsule of claim 5 wherein said UV absorbent is selected from the group consisting of salicylic acid derivatives, benzophenone derivatives and benzotriazole derivatives.

12. The microcapsule of claim 1 wherein said wall material is gelatin.

13. The microcapsule of claim 1 wherein said oily liquid substance is selected from the group consisting of a natural mineral oil, an animal oil, a plant oil and a synthetic oil.

14. The microcapsule of claim 1 wherein said wall material is selected from the group consisting of polyurethane, polyurea, polyurethane-urea, melamine and gelatin.

16. The microcapsule of claim 6 wherein said marking substance is selected from the group consisting of a fluorescent substance, a UV absorbent and a dye.

17. The microcapsule of claim 16 wherein said marking substance is said dye.

18. The microcapsule of claim 1 capable of effecting an agglutination reaction.

19. The microcapsule of claim 1 wherein said antigens or antibodies are bound to said surface by cross-linking, covalent binding or coupling.

20. The microcapsule of claim 1 wherein said antigen or antibody is bound to said surface via an aldehyde.

21. The microcapsule of claim 20 wherein said surface contains amino, imino or hydroxy groups.

22. The microcapsule of claim 1 wherein said wall material is substantially impermeable.

34. The microcapsule of claim 1 wherein said marking substance is a magnetic substance.

35. The microcapsule of claim 34 wherein said magnetic substance is selected from the group consisting of iron powders, nickel, cobalt, CrO₂, CoO, NiO, Mn₂ O₃, a magnetic zinc oxide, a magnetic iron oxide, an alloy of Al, Ni, Co or Cu, chalcogenides and ferricolloid powders.

36. The microcapsule of claim 1 wherein said wall material is a protein or a resin.

10. A method for determining a biological component having immunological activity which comprises mixing a microcapsule comprising an oily liquid core material and a wall material having bound thereto an antigen or antibody at the surface thereof with said biological component to thereby cause an immunological reaction therebetween, wherein a marking substance is incorporated into said core material.

11. The method for determining a biological component of claim 10 wherein said microcapsules have a specific gravity between 0.8 and 1.20.

15. The microcapsule of claim 10 wherein said wall material is selected from the group consisting of a polyurethane, a polyurea and a polyurethane-urea.

23. The method of claim 11 wherein said oily substance is an oil selected from the group consisting of a natural mineral oil, an animal oil, a plant oil and a synthetic oil.

24. The method of claim 10 wherein said wall material is selected from the group consisting of polyurethane, polyurea, polyurethane-urea, melamine and gelatin.

25. The method for determining a biological component of claim 24 wherein said microcapsule wall is composed of a member selected from the group consisting of a polyurethane, a polyurea and a polyurethane-urea.

26. The method of claim 10 wherein said marking substance is selected from the group consisting of a fluorescent substance, an UV absorbent and a dye.

27. The method of claim 26 wherein said marking substance is said dye.

28. The method of claim 10 wherein said immunological reaction is an agglutination reaction.

29. The method of claim 10 wherein said antigen or antibody is bound to said wall material by cross-linking, covalent bonding or coupling.

30. The method of claim 10 wherein said antigen or antibody is bound to said wall material via an aldehyde.

31. The method of claim 30 wherein said wall material contains amino, imino or hydroxy groups.

32. The method of claim 10 wherein said wall material is substantially impermeable.

33. The method of claim 10 wherein said marking substance is incorporated into said core material.

37. The method of claim 10 wherein said wall material is a protein or a resin.