Stable protein-nickel particles and methods of production and use thereof

• US 6,074,884 A
• Filed: 10/09/1997
• Issued: 06/13/2000
• Est. Priority Date: 10/09/1997
• Status: Expired due to Term

• Current Assignee: Coulter International Corp. (Danaher Corporation)
• Original Assignee: Coulter International Corp. (Danaher Corporation)
• Inventors: Maples, John A., Whitesell, James Keller, Burshteyn, Alexander, Siiman, Olavi
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): NGUYEN, BAO THUY L
• Application Number:
  US08/947,809
• Time in Patent Office: 978 Days
• Field of Search: 427/2.11, 427/2.13, 427/2.14, 427/131, 427/213.35, 427/215, 427/216, 427/337, 427/338, 428/403, 428/407, 435/5, 435/7.21, 436/518, 436/524, 436/525, 436/526, 436/529, 436/531, 436/532, 436/533, 436/534
• US Class Current: 436/518
• CPC Class Codes: G01N 2446/20 the magnetic material being present in the particle core
• CPC Class Codes: G01N 2446/90 characterised by small molecule linker used to couple immunoreagents to magnetic particles
• CPC Class Codes: G01N 33/5434 using magnetic particle immunoreagent carriers which constitute new materials per se
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

A stable protein-coated nickel particle useful in biological assays contains a nickel particle having removed from the surface thereof nickel oxide; a linker attached to said nickel particle, the linker having a free amino group; and a protein attached to said linker by covalently bonding to the free amino group. Methods of producing and using these oxide-free nickel-protein conjugates are disclosed.

Claims

1. A stable protein-coated nickel particle useful in biological assays, consisting essentially of:

(a) a nickel particle having removed from the surface thereof by ultrasonification nickel oxide;

(b) a linker attached to said nickel particle, said linker having a free amino group; and

(e) a protein attached to said linker by covalently bonding to said free amino group.

2. The particle according to claim 1, wherein said nickel particles (a) range in size from between about 3.0 to about 10.0 microns in diameter.

3. The particle according to claim 1, wherein said protein is an antibody.

4. The particle according to claim 3, wherein said antibody is selected from the group consisting of an anti-CD4 antibody and an anti-CD8 antibody.

5. The particle according to claim 3, wherein said protein is covalently bonded to the free amino groups of said linker by covalent bonding with sulfhydryl groups on 2-iminothiolane activated protein.

6. The particle according to claim 1 which is suspended in a buffer.

7. A stable protein-coated nickel particle useful in biological assays, consisting essentially of:

(a) nickel particle having removed from the surface thereof nickel oxide;

(b) an aminotrithiolate linker attached to said nickel particle, said linker having a free amino group; and

(c) a protein attached to said linker by covalently bonding to said free amino group.

8. The particle according to claim 7 wherein said aminotrithiolate linker is tris(3-mercaptopropyl)-N-glycylaminomethane of formula C₁₂ H₂₆ N₂ OS₃.

9. The particle according to claim 8, wherein said aminotrithiolate linker is covalently bonded to said nickel particles by covalent metal-sulfur bonds.