Gentamicin assay and products therefor

• US 4,305,720 A
• Filed: 08/20/1979
• Issued: 12/15/1981
• Est. Priority Date: 08/20/1979
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Bernstein, David
• Primary Examiner(s): Turk, Arnold
• Application Number:
  US06/068,165
• Time in Patent Office: 848 Days
• Field of Search: 424/12, 23/230 B, 23/915, 422/55, 422/61
• US Class Current: 436/534
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

A gentamicin protein conjugate adsorbed on a solid support, in particular, polystyrene particles, is crosslinked by a crosslinking agent to increase the stability of the supported conjugate. The particles also generally include an adsorbed protein to increase the strength of the crosslinking network. An improved conjugate is provided by the use of orosomucoid as the protein. The sensitized particles can be employed in an agglutination inhibition card test.

Claims

1. A product comprising:

a gentamicin-protein conjugate adsorbed on a solid support, said conjugate being crosslinked by a crosslinking agent to increase the stability thereof.

2. The product of claim 1 wherein the crosslinking agent is a dialdehyde.

3. An assay for gentamicin by agglutination wherein gentamicin-protein conjugate adsorbed on a solid particulate support is contacted with gentamicin antibody and sample, the improvement comprising:

said conjugate on said solid particulate support being the product of claim 1.

4. The product of claim 1 wherein the solid support is polymer particles.

5. The product of claim 4 wherein the polymer is polystyrene.

6. The product of claim 1 wherein the crosslinking agent is glutaraldehyde.

7. The product of claim 6 wherein the support is polystyrene particles.

8. The product of claim 1 wherein the surface of the solid support also has a protein adsorbed thereon, said conjugate and said protein being formed into a crosslinked network by the crosslinking agent.

9. The product of claim 8 wherein the support is polymer particles.

10. The product of claim 8 wherein the crosslinking agent is a dialdehyde.

11. The product of claim 10 wherein the protein adsorbed on the surface of the solid support is ovalbumin, the crosslinking agent is glutaraldehyde and the support is polystyrene particles.

12. A product comprising:

a conjugate of gentamicin and orosomucoid, said conjugate being supported on a solid support.

13. The product of claim 12 wherein the solid support is a particulate support.

14. In an agglutination assay for gentamicin wherein supported gentamicin is contacted with a sample and antibody to gentamicin, the improvement comprising:

said supported gentamicin being the product of claim 13.

15. The product of claim 13 wherein the conjugate is crosslinked by a crosslinking agent to increase the stability thereof.

16. The product of claim 15 wherein the crosslinking agent is glutaraldehyde.

17. The product of claim 16 wherein the solid support is polystyrene particles, said solid support having a protein adsorbed thereon, said crosslinking agent forming the protein and conjugate into a crosslinked network.

18. In an assay for gentamicin by agglutination wherein gentamicin-protein conjugate adsorbed on a solid particulate support is contacted with gentamicin antibody and sample, the improvement comprising:

said conjugate on said solid particulate support being the product of claim 16.

19. A kit for the assay of gentamicin by an agglutination procedure, comprising:

a gentamicin-protein conjugate adsorbed on a solid particulate support, said conjugate being crosslinked by a crosslinking agent to increase the stability thereof; and

antisera to gentamicin.

20. The kit of claim 19 wherein said anti-sera to gentamicin was produced by an immunogen comprised of gentamicin conjugated to a protein other than the protein of the gentamicin-protein conjugate adsorbed on the solid particulate support.

21. The kit of claim 20 wherein the protein of the supported gentamicin-protein conjugate is orosomucoid.

22. The kit of claim 21 wherein the protein of the immunogen is bovine serum albumin.

23. The kit of claim 19 and further comprising gentamicin standards.

24. The kit of claim 23 wherein the particulate support is a polymer.

25. The kit of claim 24 wherein the crosslinking agent is glutaraldehyde.

26. The kit of claim 25 wherein the surface of the solid support also has a protein adsorbed thereon, said conjugate and adsorbed protein being formed into a crosslinked network by the crosslinking agent.

27. The kit of claim 26 wherein the particles are polystyrene particles.

28. The kit of claim 27 wherein the protein of the conjugate is orosomucoid.

29. An improved card assay kit for determination of gentamicin by an agglutination procedure, comprising:

a gentamicin-protein conjugate comprised of orosomucoid as the protein supported on a solid particulate support;

an antibody to gentamicin; and

an assay card, said assay card including a plurality of separated marked areas for receiving serial dilutions of a gentamicin sample and a plurality of separated marked areas for receiving gentamicin standard, whereby serial dilution of a sample and assay thereof can be conducted on the card.

30. The kit of claim 29 and further comprising gentamicin standard.

31. The kit of claim 30 wherein the conjugate is adsorbed on the solid support, said conjugate being cross-linked by a crosslinking agent to increase the stability thereof.

32. The kit of claim 31 wherein the crosslinking agent is glutaraldehyde.

33. The kit of claim 32 wherein the particulate support is polystyrene.

34. The kit of claim 33 wherein the particulate support has a protein adsorbed on the surface thereof, said conjugate and adsorbed protein being formed in a crosslinked network by the crosslinking agent.

35. An improved card assay kit for determination of gentamicin by an agglutination procedure, comprising:

a gentamicin-protein conjugate supported on a solid particulate support;

an antibody to gentamicin;

gentamicin standard; and

an assay card, said assay card having a surface including a sample portion and a standard portion, said sample portion being comprised of at least one series of separated marked serial dilution areas for receiving at least one series of serial dilutions of a gentamicin sample, each of which has an indicia of the dilution, said standard portion including at least two separated marked standard areas for receiving gentamicin standard of different gentamicin concentration, each having an indicia of the gentamicin concentration, whereby a quantitative determination of gentamicin in a sample can be determined directly on the card by serial dilution of sample on the card, adding antibody to gentamicin and the gentamicin-protein conjugate to the serial diluted gentamicin samples and gentamicin standards and determining the lowest standard concentration and the highest dilution at which agglutination is inhibited.