Gastrin hormone immunoassays
AssigneeReceptor Biologix, Inc.
InventorsStephen Grimes, John Little, Lorraine McLoughlin
Life of this patent
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Abstract
The invention provides assay methods for the detection and quantitation of gastrin hormones, including total and free gastrin hormone in a sample. ELISA-type heterogeneous phase assays suitable for use with biological fluid samples such as blood, plasma or other bodily fluids of a mammal, particularly a human subject are provided. The method provides a precise assay for the amounts of free and total G17 and G34 in biological fluid samples, as well as the amounts of free and total Gly-extended G17, and Gly-extended G34. Also provided are methods of determining suitable treatment for patient suffering from a gastrin hormone-mediated disease or condition employing gastrin hormone immunoassays.
Claims18
A method for determining the amount of free gastrin-17 (G17) or gastrin-34 (G34) in a biological fluid sample, comprising the steps of:
- (a) obtaining a biological fluid sample comprising a gastrin hormone from a patient;
- (b) providing an immobilized antibody selected from the group consisting of an immobilized antibody that selectively binds an N-terminal epitope of G17 and an immobilized antibody that selectively binds an N-terminal epitope of G34;
- (c) incubating the sample to allow binding of G17 or G34 in the sample to said antibody to produce an immobilized complex of said antibody bound to the G17 or G34;
- (d) washing the immobilized complex to remove unbound components, and reacting the complex with a detectable marker-conjugated monoclonal antibody that selectively binds a C-terminal epitope of G17 or G34, to form a detectable marker-conjugated antibody complex;
- (e) washing the immobilized detectable marker-conjugated antibody complex, and incubating with a development reagent;
- and
- (f) measuring the developed reagent to determine the amount of free G17 or free G34 in the biological fluid sample;
- wherein the monoclonal antibody that selectively binds a C-terminal epitope of G17 or G34 is the monoclonal antibody produced by the hybridoma 458-1 (ATCC accession no. PTA-5896).
A method for determining the amount of free G34 in a biological fluid sample, comprising the steps of:
- (a) obtaining a biological fluid sample comprising the gastrin hormone G34;
- (b) providing an immobilized monoclonal antibody that selectively binds an N-terminal epitope of G34;
- (c) incubating the sample to allow binding of the G34 in the sample to said antibody to produce an immobilized complex of said antibody bound to the G34;
- (d) washing the immobilized complex to remove unbound components, and reacting the complex with a detectable marker-conjugated antibody that selectively binds a C-terminal epitope of G34, to form a detectable marker-conjugated antibody complex;
- (e) washing the immobilized detectable marker-conjugated antibody complex, and incubating with a development reagent;
- and
- (f) measuring the developed reagent to determine the amount of free G34 in the biological fluid sample;
- wherein the immobilized monoclonal antibody is the monoclonal antibody produced by the hybridoma 401-2 (ATCC accession no. PTA-5893).
A method for determining the amount of free Glycine-extended G17 or Glycine-extended G34 in a biological fluid sample, comprising the steps of:
- (a) obtaining a biological fluid sample comprising a gastrin hormone from a patient;
- (b) providing an immobilized antibody selected from the group consisting of an immobilized antibody that selectively binds an N-terminal epitope of Glycine-extended G17 and an immobilized antibody that selectively binds an N-terminal epitope of Glycine-extended G34;
- (c) incubating the sample to allow binding of Glycine-extended G17 or Glycine-extended G34 in the sample to said antibody to produce an immobilized complex of said antibody bound to the Glycine-extended G17 or Glycine-extended G34;
- (d) washing the immobilized complex to remove unbound components, and reacting the complex with a detectable marker-conjugated monoclonal antibody that selectively binds a C-terminal epitope of Glycine-extended G17 or Glycine-extended G34, to form a detectable marker-conjugated antibody complex;
- (e) washing the immobilized detectable marker-conjugated antibody complex, and incubating with a development reagent;
- and
- (f) measuring the developed reagent to determine the amount of free Glycine-extended G17 or Glycine-extended G34 in the biological fluid sample;
- wherein the monoclonal antibody is the monoclonal antibody produced by the hybridoma 445-1 (ATCC accession no. PTA-5894) or the monoclonal antibody produced by the hybridoma 445-2 (ATCC accession no. PTA-5895).
A method for determining the amount of free Glycine-extended G34 in a biological fluid sample, comprising the steps of:
- (a) obtaining a biological fluid sample comprising the gastrin hormone Glycine-extended G34;
- (b) providing an immobilized monoclonal antibody that selectively binds an N-terminal epitope of Glycine-extended G34;
- (c) incubating the sample to allow binding of the Glycine-extended G34 in the sample to said antibody to produce an immobilized complex of said antibody bound to the Glycine-extended G34;
- (d) washing the immobilized complex to remove unbound components, and reacting the complex with a detectable marker-conjugated antibody that selectively binds a C-terminal epitope of Glycine-extended G34, to form a detectable marker-conjugated antibody complex;
- (e) washing the immobilized detectable marker-conjugated antibody complex, and incubating with a development reagent;
- and
- (f) measuring the developed reagent to determine the amount of free Glycine-extended G34 in the biological fluid sample;
- wherein the immobilized monoclonal antibody is the monoclonal antibody produced by the hybridoma 401-2 (ATCC accession no. PTA-5893).
Dependencies are inferred from the claim text.