Double antibody conjugate
First Claim
1. An immunoassay reagent for detecting an antigen comprising:
- (a) a first immunoglobulin;
(b) a second immunoglobulin of different specificity than said first immunoglobulin and specific for the antigen to be detected; and
(c) a hetero-bifunctional coupling reagent selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin.
1 Assignment
0 Petitions
Accused Products
Abstract
Reagents and methods are described for an immunoassay test of increased sensitivity and decreased complexity employing an immunoglobulin specific for an antigen naturally or artificially placed upon the surface of an indicator particle coupled through the use of a hetero-bifunctional coupling reagent to a second antibody of differing specificity and specific for the antigen to be detected. In a preferred embodiment, a hetero-bifunctional coupling agent couples via a sulfhydryl group, a univalent immunoglobulin specific for the surface antigens on erythrocytes to a second multivalent immunoglobulin through an amide linkage with the latter immunoglobulin wherein said second immunoglobulin is specific for hepatitis-B surface antigen.
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Citations
23 Claims
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1. An immunoassay reagent for detecting an antigen comprising:
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(a) a first immunoglobulin; (b) a second immunoglobulin of different specificity than said first immunoglobulin and specific for the antigen to be detected; and (c) a hetero-bifunctional coupling reagent selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin.
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2. An immunoassay reagent for detecting an antigen and employing the use of an indicator substance comprising:
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(a) a first immunoglobulin specific for said indicator substance; (b) a second immunoglobulin specific for the antigen to be detected; and (c) a hetero-bifunctional coupling reagent having a sulfhydryl-reactive functional group and an amine-reactive functional group, selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin. - View Dependent Claims (3, 4, 5, 12, 13)
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6. An immunoassay reagent for detecting an antigen comprising:
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(a) an indicator substance having an associated antigen; (b) a first immunoglobulin specific for said associated antigen; (c) a second immunoglobulin specific for the antigen to be detected; and (d) a hetero-bifunctional coupling reagent selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin. - View Dependent Claims (7, 8, 9, 10, 11)
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14. A method for detecting the presence of a specified antigen within an aqueous sample comprising the steps of:
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(i) combining with said sample an immunoassay reagent for detecting an antigen comprising; (a) an indicator substance having an associated antigen; (b) a first immunoglobulin specific for said associated antigen; (c) a second immunoglobulin specific for the antigen to be detected; (d) and a hetero-bifunctional coupling reagent selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin; (ii) agitating said combination; (iii) allowing said agitated combination to settle undisturbed; and
observing said combination for the indicator substance whereby the presence of said antigen is indicated. - View Dependent Claims (16, 17, 18)
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15. A method for detecting the presence of a specified antigen within an aqueous sample comprising the steps of:
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(i) combining with said sample (a) an immunoassay reagent for detecting an antigen employing an indicator substance having an associated antigen comprising;
a first immunoglobulin specific for said associated antigen, a second immunoglobulin specific for the antigen to be detected, and a hetero-bifunctional coupling reagent selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin, and(b) said indicator substance; (ii) agitating said combination; (iii) allowing said agitated combination to settle undisturbed; and
observing said combination for the indicator substance whereby the presence of the antigen to be detected is indicated.
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19. A method for detecting the presence of a specified antigen present on a surface comprising the steps of:
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(i) contacting said surface with an immunoassay reagent employing the use of an indicator substance comprising; (a) a first immunoglobulin specific for said indicator substance; (b) a second immunoglobulin specific for said antigen to be detected; and (c) a hetero-bifunctional coupling reagent having a sulfhydryl-reactive functional group and an amine-reactive functional group, selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin; (ii) further contacting with said surface an indicator substance selected from the group consisting of erythrocytes, enzymes, radioisotopes, fluorochromes, electron opaque substances, and polymeric microspheres; (iii) allowing said reagent and said indicator substance to react; (iv) removing unreacted indicator substance; and (v) examining said surface for the presence of said indicator substance whereby the existence of said specified antigen may be determined. - View Dependent Claims (21, 22, 23)
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20. A method for detecting the presence of a specified antigen present on a surface comprising the steps of:
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(i) contacting said surface with an immunoassay reagent for detecting said specific antigen comprising; (a) an indicator substance, having an associated antigen, and selected from the group consisting of erythrocytes, enzymes, radioisotopes, fluorochromes, electron opaque substances, and polymeric microspheres; (b) a first immunoglobulin specific for said associated antigen; (c) a second immunoglobulin specific for the specified antigen to be detected; and (d) a hetero-bifunctional coupling reagent capable of selectively coupling said first immunoglobulin to at least part of the interchain disulfide linkage of said second immunoglobulin; (ii) allowing said reagent to react with said surface; (iii) removing unreacted reagent; and (iv) examining said surface for the presence of said indicator particle whereby the existence of said specified antigen may be determined.
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Specification