Homogeneous immunoassays for multiple allergens
First Claim
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1. A homogeneous immunoassay method of testing for an allergic response in a patient, comprising:
- combining an undiluted serum sample from the patient with labeled allergen-coupled particles under conditions that enable binding of the allergen-coupled particles to allergen-specific IgE antibodies from the patient'"'"'s serum sample, wherein the combining creates a response mixture;
adding a first binding agent to the response mixture, the first binding agent comprising an anti-human IgE antibody covalently bound to a first member of a binding pair;
adding a labeled second binding agent to the response mixture containing the first binding agent, the labeled second binding agent comprising a second member of the binding pair and a label, wherein the second member of the binding pair has an affinity for binding to the first member of the binding pair;
determining the amounts of the allergen-specific antibodies from the serum sample that bound to specific allergens by (i) identifying and distinguishing the labeled particles to identify subsets of specific allergens and (ii) measuring the amount of labeled second binding agent in each of the subsets of allergens;
wherein, the first binding agent and the second binding agent are preselected to enable detection of anti-human IgE antibodies at a detection sensitivity ranging from about 10−
9 g/ml to about 10−
12 g/ml, and the undiluted serum is limited to having a volume of about 1 μ
L to about 25 μ
L;
and wherein, the detection sensitivity ranging from 10−
9 g/ml to 10−
12 g/ml is measured under homogeneous assay conditions.
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Abstract
A homogeneous immunoassay method and system for quantitative determination of total immunoglobulin E and specific antibody levels to a plurality of allergens, in which a relatively small sampling of blood is required. The method utilizes relatively small microparticles in aqueous suspension. The immunoassay procedure is an immunometric sandwich procedure preferably utilizing biotin-streptavidin signal amplification techniques and R-phycoerytherin fluorescent labels.
7 Citations
6 Claims
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1. A homogeneous immunoassay method of testing for an allergic response in a patient, comprising:
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combining an undiluted serum sample from the patient with labeled allergen-coupled particles under conditions that enable binding of the allergen-coupled particles to allergen-specific IgE antibodies from the patient'"'"'s serum sample, wherein the combining creates a response mixture; adding a first binding agent to the response mixture, the first binding agent comprising an anti-human IgE antibody covalently bound to a first member of a binding pair; adding a labeled second binding agent to the response mixture containing the first binding agent, the labeled second binding agent comprising a second member of the binding pair and a label, wherein the second member of the binding pair has an affinity for binding to the first member of the binding pair; determining the amounts of the allergen-specific antibodies from the serum sample that bound to specific allergens by (i) identifying and distinguishing the labeled particles to identify subsets of specific allergens and (ii) measuring the amount of labeled second binding agent in each of the subsets of allergens; wherein, the first binding agent and the second binding agent are preselected to enable detection of anti-human IgE antibodies at a detection sensitivity ranging from about 10−
9 g/ml to about 10−
12 g/ml, and the undiluted serum is limited to having a volume of about 1 μ
L to about 25 μ
L;and wherein, the detection sensitivity ranging from 10−
9 g/ml to 10−
12 g/ml is measured under homogeneous assay conditions. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification