# US 4,391,795 A · Assay for free thyroid hormone

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- Number: US 4,391,795 A
- Document ID: US4391795A
- Status: Granted
- Kind: A
- Application number: 06/131,934
- Filed: 1980-03-21
- Granted: 1983-07-05
- Earliest claimed: 1980-03-21
- Assignees: Becton, Dickinson and Company
- Inventors: Samuel R. Pearlman
- Web page: https://patentgrep.com/patent/US4391795A

## CPC

- G01N 33/78: Thyroid gland hormones {, e.g. T3, T4, TBH, TBG or their receptors}
- Y10S 436/808: Automated or kit
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Sample containing free and bound thyroid hormone; in particular, thyroxine, is contacted with immobilized binder; in particular, supported on a solid support, to bind the free thyroxine. After separating the bound thyroxine from the immobilized binder having free thyroxine bound thereto, the immobilized binder is contacted with labeled thyroxine; in particular, radiolabeled thyroxine. Unbound labeled thyroxine is removed, and the amount of bound and/or unbound labeled thyroxine is measured, with the free thyroxine concentration in the sample being inversely proportional to the amount of labeled thyroxine bound to the immobilized binder.

## Claims

### Claim 1

A process for determining free thyroid hormone in a serum sample, comprising:
introducing a serum sample containing free thyroid hormone and thyroid hormone bound to serum proteins into a tube coated with a binder for free thyroid hormone, said binder having a number of binding sites in excess of that required to bind all of the free thyroid hormone in the serum sample, said serum sample containing the free and bound thyroid hormone being in contact with said binding sites;
incubating said serum sample in said tube to bind free thyroid hormone to said binder;
removing from the tube the serum sample which is not bound to said coated binder;
thereafter introducing into said tube labelled thyroid hormone in an amount in excess of the remaining binding sites of the coated binder to bind a portion of the labelled thyroid hormone to the coated binder;
removing from the tube unbound labelled thyroid hormone; and
measuring the amount of at least one of the bound and unbound labelled thyroid hormone for determining the amount of free thyroid hormone present in the serum sample.

### Claim 2 (depends on 1)

The process of claim 1 wherein the binder is an antibody.

### Claim 3 (depends on 2)

The process of claim 2 wherein the labeled thyroid hormone is radiolabeled.

### Claim 4 (depends on 3)

The process of claim 3 wherein the label is radioiodine.

### Claim 5 (depends on 4)

The process of claim 4 wherein the sample amount is from 50 to 200 .mu.l.

### Claim 6 (depends on 1)

The process of claim 1 wherein the thyroid hormone is T.sub.4.

### Claim 7 (depends on 6)

The process of claim 6 wherein the binder is an antibody.

### Claim 8 (depends on 7)

The process of claim 7 wherein the labeled T.sub.4 hormone is radiolabeled.

### Claim 9 (depends on 8)

The process of claim 8 wherein the label is radioiodine.

### Claim 10 (depends on 9)

The process of claim 9 wherein the sample amount is from 50 to 200 .mu.l.

### Claim 11

A kit for determining free thyroid hormone in a serum sample, comprising:
a tube coated with a binder for free thyroid hormone, said binder having a number of binding sites in excess of that required to bind all of the free thyroid hormone in a thyroid hormone standard or serum sample containing free thyroid hormone and thyroid hormone bound to serum protein which is in contact with the binding sites;
labelled thyroid hormone in an amount in excess of that required for saturating all of the binding sites after contact with the standard or serum samples; and
thyroid hormone standards of different thyroid hormone concentrations.

### Claim 12 (depends on 11)

The kit of claim 11 wherein each of the thyroid standards is provided in an amount of from 50 to 200 .mu.l.

### Claim 13 (depends on 12)

The kit of claim 12 wherein the labeled thyroid hormone is radioiodine labeled thyroid hormone.

### Claim 14 (depends on 13)

The kit of claim 13 wherein the thyroid hormone standards are provided with different thyroid hormone concentrations over a range of from 0 to 6.2 ng/dl.

### Claim 15 (depends on 11)

The kit of claim 11 wherein the thyroid hormone is T.sub.4.

### Claim 16 (depends on 13)

The kit of claim 13 wherein the thyroid hormone is T.sub.4.