Competitive binding thyroid assay with improved bound-free separation step

• US 4,015,939 A
• Filed: 05/12/1976
• Issued: 04/05/1977
• Est. Priority Date: 05/12/1976
• Status: Expired due to Term

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Lewin, Nathan, Mangiardi, Vito J.
• Primary Examiner(s): Scovronek, Joseph
• Application Number:
  US05/685,595
• Time in Patent Office: 328 Days
• Field of Search: 23/230 B, 23/230.3, 23/230.6, 210/DIG. 23, 424/1, 424/1.5, 250/303, 250/304, 260/2.1 R, 260/9
• US Class Current: 436/500
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• 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

Improved competitive binding thyroid hormone assay in which thyroid hormone bound by thyroid binding protein is separated from free thyroid hormone by removal with an intermediate base anion exchange resin. Typical are resins characterized by having an aliphatic lattice, particularly, a polyalkyleneamine lattice having tertiary and quaternary amine groups. The resin is advantageously added for the separation function in tablet form.

Claims

1. In a competitive binding assay for serum thyroid hormone having the step of separating unbound thyroid hormone from thyroid hormone bound by thyroid binding protein, the improvement comprising:

separating unbound thyroid hormone by removal with an intermediate base anion exchange resin.

2. The improved competitive binding assay in accordance with claim 1, wherein said anion exchange resin has an aliphatic lattice.

3. The improved competitive binding assay in accordance with claim 2, wherein said anion exchange resin comprises tertiary and quaternary amines on a polyalkyleneamine lattice.

4. The improved competitive binding assay in accordance with claim 3, wherein the active groups in the resin include R -- N⁺ (CH₃)₂ Cl^- and R -- N⁺ (CH₃)₂ (C₂ H₄ OH) Cl^-, and R = polyalkyleneamine lattice.

5. A clinical test for determining thyroid function comprising:

providing a test solution containing a serum sample, treating the serum sample to render the thyroid hormone in a form suitable for competitive binding, adding a thyroid hormone radioactive tracer and exogenous thyroid hormone binding protein to form a competitive binding solution and competitively bind a portion of the test serum thyroid hormone and radioactive tracer with said exogenous thyroid binding protein, contacting said competitive binding solution with intermediate base ion exchange resin, separating said resin together with free thyroid hormone and radioactive tracer that are not bound to said exogenous thyroid binding protein, and determining the magnitude of radioactivity in at least one of (1) said exogenous thyroid hormone binding protein and (2) said ion exchange resin.

6. A test for determining thyroid function in accordance with claim 5, wherein said intermediate base anion exchange resin is in tablet form upon initial contact with said competitive binding solution and is distributed therein with mixing.

7. A test for determining thyroid function n accordance with claim 6, wherein the resin in said tablet is selected to constitute about 3.3-133 mg for each 0.1 ml of serum in the test solution.

8. A test for determining thyroid function in accordance with claim 6, wherein the resin in said tablet is selected to constitute about 8.3-66.7 mg for each 0.1 ml of serum in the test solution.

9. A test in accordance with claim 5, wherein said resin is about 50-400 wet mesh.

10. A test in accordance with claim 9, wherein the resin is about 200-400 wet mesh.