Homogeneous chemiluminescence immunoassay using a light absorbing material
First Claim
1. A method for determining an unknown quantity of an analyte in a fluid comprising:
- (a) preparing a mixture comprising an antianalyte attached to the surface of a tube, an analyte in a fluid, a light absorbing material, and a tracer for said analyte, said tracer including a chemiluminescent label;
(b) causing binding involving immunologically specific pairs of said antianalyte, said analyte and said tracer whereby bound tracer is formed on the surface of said tube;
(c) adding to said mixture a composite comprising an activator of a chemiluminescence source whereby chemiluminescence is induced;
(d) measuring chemiluminescence associated with said bound tracer; and
(e) determining the quantity of said analyte in said fluid by comparing the magnitude of said chemiluminescence with the magnitude of chemiluminescence established for a known quantity of the analyte.
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Abstract
A method for separation-free solid phase immunoassay of an analyte includes contacting an antianalyte attached to the surface of a solid support with the analyte, a light absorbing material and a tracer for the analyte which includes a label. The resulting mixture is incubated and chemiluminescence is generated. All of the chemiluminescence is absorbed by the light absorbing material except that associated with the bound tracer whereby the only emission detected is due to the bound tracer. Since emission from free tracer in the fluid phase of the assay medium is not detected, separation of the bound and free fractions is unnecessary. The invention includes a kit of materials useful in performing an immunoassay in accordance with the method of the invention.
29 Citations
18 Claims
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1. A method for determining an unknown quantity of an analyte in a fluid comprising:
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(a) preparing a mixture comprising an antianalyte attached to the surface of a tube, an analyte in a fluid, a light absorbing material, and a tracer for said analyte, said tracer including a chemiluminescent label; (b) causing binding involving immunologically specific pairs of said antianalyte, said analyte and said tracer whereby bound tracer is formed on the surface of said tube; (c) adding to said mixture a composite comprising an activator of a chemiluminescence source whereby chemiluminescence is induced; (d) measuring chemiluminescence associated with said bound tracer; and (e) determining the quantity of said analyte in said fluid by comparing the magnitude of said chemiluminescence with the magnitude of chemiluminescence established for a known quantity of the analyte. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method for determining an unknown quantity of an analyte in a fluid comprising:
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(a) preparing a mixture comprising an antianalyte attached to the surface of a tube, an analyte in a fluid, a light absorbing material and a tracer which includes a peroxidase; (b) causing binding involving immunologically specific pairs of said antianalyte, said analyte and said tracer whereby bound tracer is formed on the surface of said tube; (c) adding to said mixture a substrate for said peroxidase said substrate reacting with said peroxidase to cause chemiluminescence wherein substantially all of said chemiluminescence is absorbed by said light absorbing material except that caused by reaction of said substrate with said bound tracer; measuring chemiluminescence associated with bound tracer; and (e) determining the quantity of said analyte in said fluid by comparing the magnitude of said chemiluminescence with the magnitude of chemiluminescence established for a known quantity of the analyte.
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13. A kit of materials for performing a chemiluminescence immunoassay for an unknown quantity of an analyte in a fluid comprising:
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(a) a tube having attached thereto an antianalyte specific to an analyte; (b) a light absorbing material; and (c) a tracer for said analyte, said tracer comprising a label, said label being a first component of a chemiluminescence source. - View Dependent Claims (14, 15, 16, 17, 18)
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Specification