US 5,210,017 A · Ligand-mediated immunofunctional hormone binding protein assay method

Abstract

A ligand-mediated immunofunctional assay (LIFA) method for detecting the presence and the concentration of polypeptide hormone binding proteins which comprises capturing the binding protein with a solid phase bound first antibody, saturating the bound hormone binding protein with the ligand polypeptide hormone, and detecting the bound ligand polypeptide hormone with a detectably labeled second antibody specific for the ligand polypeptide hormone. In the absence of added saturating polypeptide hormone, the LIFA measures the amount of hormone binding protein bound to the endogenous ligand polypeptide hormone. A growth hormone binding protein assay illustrates the method of the present invention.

Claims24

Dependent claims

A ligand-mediated immunofunctional method for determining the amount of polypeptide hormone binding protein in a liquid sample comprising:

(a) contacting said liquid sample with:
1) a first antibody attached to a solid phase carrier, wherein said first antibody is specific for epitopes on said polypeptide hormone binding protein such that in the presence of antibody the hormone binding sites remain available on the binding protein for binding to the ligand polypeptide hormone, thereby forming a complex between said first antibody and said polypeptide hormone binding protein;
and 2) said ligand polypeptide hormone for a period of time sufficient to saturate all available ligand hormone binding sites on said hormone binding protein thereby forming a saturated complex;
(b) contacting said saturated complex with a detectably labeled second antibody which is specific for epitopes on said ligand polypeptide hormone which are available for binding when said ligand polypeptide hormone is bound to said hormone binding protein;
and
(c) quantitatively analyzing the amount of said second labeled antibody bound as a measure of said polypeptide hormone binding protein in said liquid sample.

3 more elements

A method for determining the amount of endogenous ligand polypeptide hormone bound to a polypeptide hormone binding protein in a liquid sample comprising:

(a) contacting said liquid sample with a first antibody attached to a solid phase carrier, wherein said first antibody is specific for epitopes on said polypeptide hormone binding protein such that in the presence of antibody the hormone binding sites remain available on the binding protein for binding to a ligand polypeptide hormone, thereby forming a complex between said first antibody and said polypeptide hormone binding protein;
(b) contacting said complex with a detectably labeled second antibody which is specific for epitopes on said ligand polypeptide hormone which are available for binding when said ligand polypeptide hormone is bound to said hormone binding protein;
and
(c) determining the amount of said second labeled antibody bound as a measure of endogenous polypeptide hormone bound to said polypeptide hormone binding protein in said liquid sample.

1 more element

Dependencies are inferred from the claim text.

Ligand-mediated immunofunctional hormone binding protein assay method

Life of this patent

Ligand-mediated immunofunctional hormone binding protein assay method

Life of this patent

Abstract

Claims24