Immobilized metal colloids on dispersed polymer microspheres

• US 5,552,086 A
• Filed: 09/09/1993
• Issued: 09/03/1996
• Est. Priority Date: 01/29/1992
• Status: Expired due to Fees

• Current Assignee: Coulter International Corp.
• Original Assignee: Coulter Corporation (Danaher Corporation)
• Inventors: Cayer, Marilyn, Burshteyn, Alexander, Siiman, Olavi
• Primary Examiner(s): Lovering, Richard D.
• Application Number:
  US08/118,980
• Time in Patent Office: 1,090 Days
• Field of Search: 252/315.2, 252/408.1, 428/402.24, 428/403, 428/407, 428/461, 428/493, 427/2.23, 427/213.31, 427/214, 436/170, 424/9.1
• US Class Current: 252/408.1
• CPC Class Codes: A61K 47/6923 the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
• CPC Class Codes: A61K 47/6929 the form being a nanoparticle, e.g. an immuno-nanoparticle
• CPC Class Codes: G01N 33/548 Carbohydrates, e.g. dextran
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: Y10T 428/2989 Microcapsule with solid core [includes liposome]
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

The invention describes stable colloidal polymeric particles of 0.2-5.0 microns size which are coated with a first aminodextran layer and a second metallic solid layer, The metal is coated by reduction of a metallic salt or complex by the aminodextran. The metal coated particles, which preferably are gold- or silver-coated polymeric microspheres, produce side scatter and forward shifts in flow cytometry applications that are in agreement with the theoretical shifts predicted for solid gold or silver spheres of similar size in flow cytometry applications,

Claims

1. Stable colloidal particles suitable for use in instrumental analyses such as flow cytometry or Raman spectroscopy, each such particle comprised of a colloidal-sized core substrate, an aminodextran coating over the peripheral surface of said substrate and a uniform layer of colloidal-sized metallic solid overlaying said aminodextran coating, wherein the metal for said metallic solid is selected from the group consisting of metals which can be reduced from the ionic state to the metal (O) state by the aminodextran coating the core substrate.

2. The particles of claim 1 wherein the metallic solid is selected from the group consisting of metals which form metal ions or metal ion complexes which have a reduction potential of +0.7 volts or higher.

3. The particles of claim 2 wherein said metal is selected from the group consisting of silver and gold.

4. The particles of claim 3 wherein said aminodextran is selected from the group consisting of aminodextrans in which the degree of diamine substitution in said aminodextran is in the range of 1/20- 1/5 compared to a maximum theoretical value of 1/2.5.

5. The particles of claim 4 wherein the diamine substitution in said aminodextran is approximately 1/6.

6. The particles of claim 5 wherein the colloidal core substrate is polymeric.

7. The particles of claim 6 wherein the polymer core substrate is polystyrene.

8. The particles of claim 7 wherein the polystyrene core substrate is in the size range 0.2-5.0 microns.

9. The particles of claim 8 wherein the polystyrene core substrate is in the size range 0.2-2.2 microns.

10. The particles of claim 1 wherein said aminodextran is selected from the group consisting of aminodextrans in which the degree of diamine substitution in said aminodextran is in the range of 1/20- 1/5 compared to a maximum theoretical value of 1/2.5.

11. The particles of claim 10 wherein the diamine substitution in said aminodextran is approximately 1/6.

12. The particles of claim 1 wherein said colloidal sized core substrate is a polymeric substrate.

13. The particles of claim 12 wherein the polymer core substrate is polystyrene.

14. The particles of claim 13 wherein the polystyrene core substrate is in the size range 0.2-5.0 microns.

15. The particles of claim 14 wherein the polystyrene core substrate is in the size range 0.2-2.2 microns.

16. Stable colloidal particles suitable for use in instrumental analyses such as flow cytometry or Raman spectroscopy, each such particle comprised of a colloidal-sized metallic solid deposited on the peripheral surface of an aminodextran-coated, colloidal-sized core polymeric substrate;

said aminodextran being covalently bonded to said core polymeric substrate by a covalent bond between aminodextran amine groups and amine-reactive functional groups on said core polymeric substrate; and

said metallic solid and said aminodextran being uniformly dispersed on said peripheral surface, wherein the metal for said metallic solid is selected from the group consisting of metals which can be reduced from the ionic state to the metal (O) state by the aminodextran coating the polymeric core substrate.

17. The particles of claim 16 wherein the metallic solid is selected from the group consisting of metals which form metal ions or metal ion complexes which have a reduction potential of +0.7 volts or higher.

18. The particles of claim 17 wherein the metallic solid is selected from the group consisting of silver and gold.

19. The particles of claim 18 wherein said aminodextran is selected from the group consisting of aminodextrans in which the degree of diamine substitution in said aminodextran is in the range of 1/20- 1/5 compared to a maximum theoretical value of 1/2.5.

20. The particles of claim 19 wherein the diamine substitution in said aminodextran is approximately 1/6.

21. The particles of claim 20 wherein the polymeric colloidal core substrate is polystyrene.

22. The particles of claim 21 wherein the polystyrene core substrate is in the size range 0.2-5.0 microns.

23. The particles of claim 22 wherein the polystyrene core substrate is in the size range 0.2-2.2 microns.

24. The particles of claim 16 wherein said aminodextran is selected from the group consisting of aminodextrans in which the degree of diamine substitution in said aminodextran is in the range of 1/20- 1/5 compared to a maximum theoretical value of 1/2.5.

25. The particles of claim 24 wherein the degree of diamine substitution in said aminodextran is about 1/6.

26. The particles of claim 16 wherein the core polymeric substrate is polystyrene.

27. The particles of claim 26 wherein the polystyrene core is in the size range 0.2-5.0 microns.

28. The particles of claim 27 wherein the polystyrene core is in the size range 0.2-2.2 microns.

29. A method of preparing polymeric particles with a uniform coating of a metallic solid on the peripheral surface of said polymeric particles, said method comprising:

(a)(1) adsorbing an aminodextran on the surface of a polymeric colloidal-sized particles, said aminodextran being capable of reducing the metal ion or metal ion complex to the metal (O) state;

or

(2) covalently binding the aminodextran of (a)(1) to the surface of a polymeric particles;

(b)(1) crosslinking the aminodextran coating of the particles of step (a)(1) by addition of a selected crosslinking agent, or

(2) crosslinking the aminodextran coating of the particles of step (a)(2) by the addition of a selected crosslinking agent or optionally not crosslinking said coating;

(c) washing the crosslinked particles of step (b);

(d) mixing together, for a selected reaction time and at a selected reaction temperature, a suspension of the particles of step (c) and a solution of a metal ion or metal ion complex capable of being reduced by the aminodextran coating on the particles of step (c) the reaction time and temperature being sufficient to allow the metal ion or metal ion complex to become coordinated to said aminodextran and subsequently reduced by said aminodextran to the metal (O) state; and

(e) separating the metal-coated particles of step (d) and washing the same before using said metal-coated particles.

30. The method of claim 29 wherein said metal ions or metal ion complexes have a reduction potential of +0.7 volts or higher.

31. The method of claim 30 wherein said metal ions or metal ion complexes are selected from the group consisting of silver and gold.

32. The method of claim 29 wherein said polymeric particles are polystyrene microbeads of diameter in the size range 0.2-5.0 microns.

33. The method of claim 32 wherein said polystyrene microspheres are in the size range 0.2-2.2 microns.

34. The method of claim 29 wherein when said metallic solid is silver or gold, said reaction time is in the range of 5 seconds to 10 minutes.

35. The method of claim 29 wherein when said metallic solid is silver or gold, said reaction temperature is in the range of 80°

-100°

C.