Assay method and kit

• US 4,311,685 A
• Filed: 06/18/1979
• Issued: 01/19/1982
• Est. Priority Date: 09/08/1978
• Status: Expired due to Term

• Current Assignee: Burroughs Wellcome and Co.
• Original Assignee: Burroughs Wellcome and Co.
• Inventors: Snyder, Solomon H.
• Primary Examiner(s): Nucker, Christine M.
• Application Number:
  US06/049,418
• Time in Patent Office: 946 Days
• Field of Search: 424/1, 424/12, 23/230 B, 422/67
• US Class Current: 424/1.85
• CPC Class Codes: G01N 33/60 involving radioactive labelled substances
• CPC Class Codes: G01N 33/9453 Cardioregulators, e.g. antihypotensives, antiarrhythmics

Abstract

Method and materials for determining levels of β-adrenergic blocking drugs in body fluid, the method including the step of measuring inhibition of the binding of β-adrenergic receptor binder to β-adrenergic receptor material caused by β-adrenergic blocker present in the body fluid.

Claims

1. The method of determining the concentration of free β

-adrenergic blocking drugs and any active metabolites thereof in a body fluid containing same comprising (a) dialyzing the body fluid (b) mixing together β

-adrenergic receptor material, radioactive β

-adrenergic receptor binder and the dialyzed body fluid, and measuring the amount of the radioactive β

-adrenergic binder on the β

-adrenergic receptor material and (c) mixing together a concentration of a standard amount of non-radioactive β

-adrenergic receptor binder, β

-adrenergic receptor material and radioactive β

-adrenergic receptor binder and measuring the amount of radioactive β

-adrenergic receptor binder on the β

-adrenergic receptor material.

2. The method of claim 1 in which the material, binder, and dialyzed body fluid are permitted to remain together a time sufficient to produce sufficient binding of the binder, drugs and any active metabolites thereof in the body fluid to said receptor material prior to making the measurement.

3. The method of claim 1 in which the material, binder and dialyzed body fluid are combined in the presence of sufficient buffer to produce a pH of about 6 to 9.

4. The method of claim 1 in which unbound binder and dialyzed body fluid are removed as part of the measurement.

5. The method of claim 1, 2, 3 or 4 in which the dialyzed body fluid is blood plasma or blood serum.

6. The method of claim 1 in which measuring of the amount of radioactive receptor binder on the receptor material is determined in a gamma detector or scintillation counter based upon the nature of the radionuclide portion of the radioactive binder.

7. The method of claim 1 or 6 in which the concentration of drug and active metabolite is determined by reference to a standard curve representing percent inhibition of radioactive receptor binder vs. non-radioactive receptor binder.

8. The method of claim 1 in which the concentration of body fluid is less than about 10%.

9. The method of claim 1 or 8 in which the dialyzed body fluid is blood plasma or blood serum.

10. The method of claim 1 or 8 in which the amount of dialyzed body fluid in the mixture containing same is greater than one microliter.

11. The method of claim 1 in which the unbound drug, unbound radioactive binder and dialyzed body fluid are removed in determining the percent inhibition of binding.

12. The method of claim 1 in which the radioactive receptor binder is a radioactive labeled drug selected from the group consisting of:

1. propranolol

2 . practalol

3. pindalol

4. alprenolol

5. sotalol

6. butoxamine

13. The method of claim 1 in which the receptor material is brain tissue.

14. The method of claim 1 in which (c) is repeated a sufficient number of times while varying concentrations of the non-radioactive binder to provide information for generating a standard curve.

15. The method of claim 9 in which the amount of dialyzed body fluid in the mixture containing same is greater than one microliter.

16. The method of claim 9 in which the unbound drug, unbound radioactive binder and dialyzed body fluid are removed in determining the percent inhibition of binding.

17. The method of claim 9 in which the radioactive receptor binder is a radioactive labeled drug selected from the group consisting of:

1. propranolol

2. practalol

3. pindalol

4. alprenolol

5. sotalol

6. butoxamine

18. The method of claim 9 in which the receptor material is brain tissue.

19. The method of claim 9 in which (c) is repeated a sufficient number of times while varying concentrations of the non-radioactive binder to provide information for generating a standard curve.