Biochemical avidin-biotin multiple-layer system

• US 4,282,287 A
• Filed: 01/24/1980
• Issued: 08/04/1981
• Est. Priority Date: 01/24/1980
• Status: Expired

• Current Assignee: Roger W Giese
• Original Assignee: Roger W Giese
• Inventors: Giese, Roger W.
• Primary Examiner(s): Smith, Ronald H.
• Assistant Examiner(s): Childs, S. L.
• Application Number:
  US06/114,898
• Time in Patent Office: 558 Days
• Field of Search: 427/2, 427/214, 427/399, 427/220, 427/400, 427/222, 427/414, 427/352, 427/331, 435/7, 435/188, 422/57, 428/403, 428/407, 428/478.2
• US Class Current: 428/407
• CPC Class Codes: A61L 2420/08 Coatings comprising two or more layers
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 33/0005 Use of materials characterised by their function or physical properties
• CPC Class Codes: B01D 67/00933 by addition of a layer chemically bonded to the membrane
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C08L 89/00 Compositions of proteins; Compositions of derivatives thereof foodstuff preparations A23J3/00
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer
• CPC Class Codes: Y10T 428/31768 Natural source-type polyamide [e.g., casein, gelatin, etc.]

Abstract

A multiple-layer process for applying, in alternate, successive layers, the protein, avidin, and a biotin-containing extender material to a solid surface to modify the properties of the surface and to the multiple-layer product so prepared.

Claims

1. A process of modifying the surface properties of a surface, which process comprises:

applying alternative, monomolecular, successive layers of first and and second materials to a surface to be modified, the first material comprising avidin and the second material comprising a noncovalent, biotin-modified extender, one of the materials reacted to the surface, and, thereafter, at least one additional layer of each of the first and second materials alternated, secured and reacted to the underlying layer, to provide a surface with the first or second material as the top surface layer thereon.

2. The process of claim 1 wherein the top surface of the process comprises the second extender material.

3. The process of claim 1 wherein the first material comprises a modified avidin adapted to react with the second extender material through the avidin portion of the modified material.

4. The process of claim 1 which includes pretreating the surface with a monomolecular layer of biotin, and wherein the first avidin material is applied over and secured to the biotin layer.

5. The process of claim 1 which includes:

(a) applying a monomolecular layer of biotin and covalently binding the biotin to the surface;

(b) applying and reacting a monomolecular layer of avidin to the biotin layer; and

(c) applying and reacting a monomolecular layer of a biotin extender material to the avidin layer.

6. The process of claim 5 which includes:

(a) applying another layer of avidin; and

(b) applying another layer of the biotin extender material to the other layer of avidin.

7. The process of claim 1, which process includes varying the concentration of the first or second material in the alternate, successive layers.

8. The process of claim 7 which includes increasing the concentration of the alternate, successive layers of the first and second materials.

9. The process of claim 7 which includes decreasing the concentration of the alternate, successive layers of the first and second materials.

10. The process of claim 1 which includes applying alternate, successive layers of approximately the same stochiometric concentration.

11. The process of claim 1 wherein the surface comprises a polymeric surface.

12. The process of claim 11 wherein the surface comprises the surface of finely-divided, polyacrylamide, polymer particles.

13. The process of claim 11 wherein the surface comprises erythrocytes.

14. The process of claim 1 wherein the surface comprises the surface of amino polyacrylamide particles, and the process comprises applying a layer of biotin-NHS esters to the surface of the particles, and, thereafter, applying successive, alternate, monomolecular layers of avidin and biotin-ribonuclease material.

15. The process of claim 1 wherein the first and second materials comprise three or more layers.

16. The process of claim 1 wherein the first and second materials comprise two monomolecular layers.

17. The process of claim 1 wherein each layer of the materials is monomolecular or monoparticulate in thickness.

18. The process of claim 1 which includes reacting at least one of the avidin layers with a biotin-horse radish peroxidase or a biotin ribonuclease as a signal extender.

19. The layering system produced by the process of claim 1.

20. The layering system produced by the process of claim 14.

21. A monomolecular-layering process of modifying the surface properties of a substrate surface of a polymer, which process comprises:

(a) applying a layer of a biotin-N-hydroxysuccinimide ester as a biotin-extender material to the surface of a polymer, to react the biotin-N-hydroxysuccinimide to the surface of the polymer;

(b) washing the polymer surface to remove unreacted biotin-N-hydroxysuccinimide;

(c) applying a layer of avidin to the washed polymer surface, to react the avidin with the biotin-N-hydroxysuccinimide extender material;

(d) washing the reacted surface to remove unreacted avidin;

(e) applying to the washed avidin surface a layer of a caproylamidobiotin NHS or RNase as a biotin-extender material;

(f) washing the reacted surface to remove unreacted biotin-extender material; and

(g) recovering the polymer having multiple layers, with the top layer composed of a biotin-extender material.

22. The process of claim 21 which includes:

(a) applying avidin to the washed polymer surface, to react another layer of avidin with the biotin-extender material;

(b) washing the reacted surface to remove unreacted avidin; and

(c) recovering a polymer having multiple layers, with the top layer composed of avidin.

23. The process of claim 22 which includes repeating the successive application of biotin-extender material and avidin, with intermittent wash steps, to provide a polymer surface with successive monomolecular layers of avidin and biotinextender material, with the top monomolecular layer being either avidin or a biotin-extender material.

24. The process of claim 21 wherein the polymer comprises an aminoalkyl polyacrylamide polymer.

25. The proces of claim 21 which includes reacting a small amount of a biotin-peroxidase or -ribonuclease material, as a signal extender for the avidin, with at least one of the applications of the avidin.

26. The polymeric-layering system produced by the process of claim 21.

27. The polymeric-layering system produced by the process of claim 22.

28. The polymeric-layering system produced by the process of claim 23.