Immobilized hydrophobically-modified antibodies

• US 5,314,830 A
• Filed: 10/30/1992
• Issued: 05/24/1994
• Est. Priority Date: 10/30/1992
• Status: Expired due to Fees

• Current Assignee: The University of New Mexico
• Original Assignee: The University of New Mexico
• Inventors: Anderson, William L., Otero, Dennis
• Primary Examiner(s): Elliott, George C.
• Application Number:
  US07/968,076
• Time in Patent Office: 571 Days
• Field of Search: 530/391.1, 530/391.9, 436/532, 436/547, 436/524, 436/527, 435/7.1, 435/181
• US Class Current: 436/524
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/552 Glass or silica

Abstract

The invention comprises stable, antigen-sensitive surfaces containing immobilized antibody modified with a polyethylene glycol spacer arm including a terminal hydrophobic moiety. The surfaces are generally useful in immunoassay procedures, especially microgravimetric assays such as those employing antibody immobilized on piezoelectric crystal surfaces.

Claims

1. A protein modified with a hydrophobic moiety attached to the protein by a spacer comprising a water-soluble polymer, directly adsorbed onto an unprimed microporous mineral body surface.

2. The modified protein of claim 1, wherein the protein is an antibody or antigen.

3. The modified protein of claim 2, wherein the water-soluble polymer is polyethylene glycol or a derivative thereof.

4. The antibody or antigen of claim 3, wherein the hydrophobic moiety is a fatty acid residue.

5. The antibody or antigen of claim 4, wherein the fatty acid residue contains from 16 to 18 carbon atoms.

6. The antibody or antigen of claim 4, wherein the hydrophobic moiety is stearyl.

7. The antibody or antigen of claim 2, wherein the surface is a piezoelectric crystal surface.

8. The antibody or antigen of claim 7, wherein the water-soluble polymer is polyethylene glycyol or a derivative thereof.

9. The antibody or antigen of claim 7, wherein the hydrophobic moiety is a fatty acid residue.

10. The antibody or antigen of claim 7, wherein the hydrophobic moiety is a fatty acid residue and the water-soluble polymer is polyethylene glycol or a derivative thereof.

11. The antibody or antigen of claim 10, wherein the polyethylene glycol has a molecular weight of from about 50 to 150.

12. The antibody or antigen of claim 11, wherein the polyethylene glycol has a molecular weight of from about 80 to 90.

13. The antibody or antigen of claim 11, wherein the polymer has a molecular weight from about 80 to 90.

14. The modified protein of claim 1, wherein the surface is a glass bead surface, a silica crystal surface, a ceramic surface, an optical fiber surface or a quartz surface.

15. The modified protein of claim 14, wherein the surface is a piezoelectric quartz crystal.

16. The modified protein of claim 15, wherein the protein is an antibody or antigen.

17. The modified protein of claim 16, wherein the water-soluble polymer is polyethylene glycol or a derivative thereof.

18. The antibody or antigen of claim 17, wherein the hydrophobic moiety is a fatty acid residue.

19. The modified protein of claim 18, wherein from about 15 to about 25% of the lysine residues of the antibody are substituted with polyethylene glycol or a derivative thereof.

20. The modified protein of claim 18, wherein the protein is adsorbed upon the surface by application to the surface followed by air drying.

21. A method for preparing an immunoreactive surface, comprising coating an unprimed microporous mineral body surface with an antigen or antibody modified with a hydrophobic moiety attached to the antigen or antibody by a spacer comprising a water-soluble polymer by drying the modified antigen or antibody onto the unprimed surface.

22. The method of claim 21, wherein the microporous mineral body surface is a piezoelectric crystal surface.

23. The method of claim 22, wherein the water-soluble polymer is polyethylene glycol having a molecular weight of from about 50 to 150.

24. The method of claim 23, wherein the hydrophobic moiety is a fatty acid residue.