Radioimmunoassay of plasma steroids

• US 4,081,525 A
• Filed: 08/16/1976
• Issued: 03/28/1978
• Est. Priority Date: 08/16/1976
• Status: Expired due to Term

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Inventors: Knight, William S., Kelley, Barbara J.
• Primary Examiner(s): Padgett, Benjamin R.
• Assistant Examiner(s): Nucker, Christine M.
• Application Number:
  US05/710,499
• Time in Patent Office: 589 Days
• Field of Search: 23/230 B, 424/1, 424/12, 424/1.5
• US Class Current: 436/538
• CPC Class Codes: G01N 33/743 Steroid hormones
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/817 Steroids or hormones
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives

Abstract

An in vitro radioimmunoassay procedure for measuring an amount of steroid in a sample wherein a separate serum protein inactivation step is eliminated. The separate protein serum inactivation step is eliminated by employing a solution wherein the solvent comprises from about 0 to about 10% of a water miscible organic solvent and from about 90 to about 100% of a buffer having a pH of from about 3 to about 6.

Claims

1. An in vitro radioimmunoassay procedure comprising:

(a) contacting a sample comprising serum proteins and steroids with a solution comprising a solvent and a solute, said solute comprising labeled steroids and specific binders for said steroids, and said solvent comprising from about 0 to about 10% by weight of a water miscible organic solvent and from about 90 to about 100% of a buffer having a pH of from about 3 to about 6, whereby the serum proteins are simultaneously inactivated while labeled and unlabeled steroids compete for binding sites on said steroid specific binders;

(b) separating bound steroids from unbound steroids into two fractions; and

(c) measuring one of said fractions for the presence of labeled steroids.

2. The procedure of claim 1 wherein said labeled steroids are labeled cortisol.

3. The procedure of claim 1 wherein said solvent comprises from about 3 to about 7 percent of said water miscible organic solvent and from about 93 to about 97 percent of said buffer having a pH of from about 4 to about 5.

4. The procedure of claim 3 wherein said labeled steroids are labeled cortisol.

5. The procedure of claim 1 wherein said sample and solution are present in amounts such that said sample has a final sample solution of from about 1/10 to about 1/100.

6. The procedure of claim 5 wherein said labeled steroids are labeled cortisol.

7. The procedure of claim 5 wherein said solvent comprises from about 3 to about 7 percent of said water miscible organic solvent and from about 93 to about 97 percent of said buffer having a pH of from about 4 to about 5.

8. The procedure of claim 7 wherein said labeled steroids are labeled cortisol.

9. The procedure of claim 7 wherein said organic solvent is selected from a group consisting of methanol, ethanol, dioxane, acetone, dimethyl formamide, ehtylene glycol and propylene glycol, and wherein said buffer is selected from a group consisting of a metal salt of acetic acid, a metal salt of citric acid, a metal salt of maleic acid, glutamate-acetate, glutamate-citrate, glutamate-hydrogen chloride, glycinate-hydrogen chloride, and glycinate-phosphate.

10. The procedure of claim 9 wherein said labeled steroids are labeled cortisol.

11. The procedure of claim 9 wherein said organic solvent is selected from the group consisting of methanol and ethanol and wherein said buffer is selected from a group consisting of sodium citrate and sodium acetate.

12. The procedure of claim 11 wherein said labeled steroids are labeled cortisol.

13. The procedure of claim 11 wherein said solvent comprises about 5% of said organic solvent and about 95% of said buffer.

14. The procedure of claim 13 wherein said labeled steroids are labeled cortisol.

15. The procedure of claim 13 wherein said sample and solution are present in amounts such that said sample has a final sample dilution of from about 1/25 to about 1/50.

16. The procedure of claim 15 wherein said labeled steroids are labeled cortisol.

17. The procedure of claim 1 wherein said solvent comprises from about 0 to about 3 percent of said water miscible organic solvent and from about 97 to about 100 percent of said buffer having a pH of from about 3 to about 5.

18. The procedure of claim 17 wherein said labeled steroids are labeled cortisol.

19. The procedure of claim 17 wherein said sample and solution are present in amounts such that said sample has a final sample dilution of from about 1/10 to about 1/100.

20. The procedure of claim 19 wherein said labeled steroids are labeled cortisol.