US4278653A
Expired / Lapsed
US
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Methods and kits for double antibody immunoassay providing a colored pellet for easy visualization
Assignee: Revvity Health Sciences Inc
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Filing: 1979-02-01
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Granted: 1981-07-14
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Expires: 1999-02-01
Abstract
Abstract Methods and kits for performing a double antibody immunoassay are described. The immunoassay comprises: reacting an antigen and a labelled version of said antigen with a first antibody against said antigen and said first antibody with a second antibody against said first antibody to precipitate a complex of said antigen, said first antibody and said second antibody, and precipitating a colored protein to form a precipitate comprised of said complex and colored protein. In a preferred embodiment, the first and second antibodies can be prereacted to simplify the assay procedure.
Inventors
- 👤 S. Richard Harris
- 👤 Darlene K. Bonci
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 21:13
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/807; Y10S436/808; Y10S436/815; Y10S436/817
Claims Analysis
Total Claims: 58
Claim 1
Independent Claim
68 words
A method for performing an immunoassay, said method comprising: reacting an antigen and a labelled version of said antigen with a first antibody for said antigen and said first antibody with a second antibody for said first antibody to precipitate a complex of said antigen, said first antibody and said second antibody, and precipitating a colored protein to form a precipitate comprised of said complex and colored protein.
Claim 2
Independent Claim
68 words
A method for performing an immunoassay, said method comprising: reacting an antigen and a labelled version of said antigen with a first antibody for said antigen and said first antibody with a second antibody for said first antibody to precipitate a complex of said antigen, said first antibody and said second antibody, and precipitating a colored protein to form a precipitate comprised of said complex and colored protein.
Claim 3
Dependent Claim
40 words
The method of claim 1 wherein said precipitation of colored protein is effected by adding a precipitating agent in a sufficient amount to precipitate at least a portion of the colored protein to form a colored precipitate for easy visualization.
Claim 4
Dependent Claim
21 words
The method of claim 1 wherein said antigen is selected from the group consisting of gentamicin, tobramycin, theophylline, digoxin, and hydrocortisone.
Claim 5
Dependent Claim
23 words
The method of claim 1 wherein said colored protein is formed by mixing a protein and a protein dye in an aqueous solution.
Claim 6
Dependent Claim
18 words
The method of claim 3 wherein said protein has an average molecular weight of at least about 100,000.
Claim 7
Dependent Claim
25 words
The method of claim 3 wherein said dye is present in the range of about 0.0001 to 0.1% by weight of the final assay mixture.
Claim 8
Dependent Claim
24 words
The method of claim 3 wherein said protein is present in an amount of at least 0.1% by weight of the final assay mixture.
Claim 9
Dependent Claim
24 words
The method of claim 1 wherein said precipitating agent is selected from the group consisting of polyethylene glycol, polylysine, dextran, polyvinylpyrolidone and polyvinyl alcohol.
Claim 10
Dependent Claim
22 words
The method of claim 1 wherein said precipitating agent is polyethylene glycol having an average molecular weight of at least about 6000.
Claim 11
Dependent Claim
26 words
The method of claim 1 wherein said precipitating agent is present in an amount of at least about 11/2% by weight of the final assay mixture.
Claim 12
Dependent Claim
14 words
The method of claim 1 wherein said labelled version of the antigen is radio-labelled.
Claim 13
Independent Claim
108 words
A method for performing an immunoassay, said method comprising: mixing an aqueous mixture of (1) a sample containing an antigen to be assayed, (2) a known quantity of a labelled version of the antigen, and (3) a first antibody for the antigen in an amount insufficient to react with all the antigen, thus forming a first antibody-antigen complex; and adding to said aqueous mixture (4) a second antibody that reacts with the first antibody-antigen complex to form a precipitate, (5) a colored protein and (6) a precipitating agent that effects the precipitation of at least some of the colored protein; thus forming a colored precipitate for easy visualization.
Claim 14
Independent Claim
108 words
A method for performing an immunoassay, said method comprising: mixing an aqueous mixture of (1) a sample containing an antigen to be assayed, (2) a known quantity of a labelled version of the antigen, and (3) a first antibody for the antigen in an amount insufficient to react with all the antigen, thus forming a first antibody-antigen complex; and adding to said aqueous mixture (4) a second antibody that reacts with the first antibody-antigen complex to form a precipitate, (5) a colored protein and (6) a precipitating agent that effects the precipitation of at least some of the colored protein; thus forming a colored precipitate for easy visualization.
Claim 15
Dependent Claim
21 words
The method of claim 12 wherein said antigen is selected from the group consisting of gentamicin, tobramycin, theophylline, digoxin and hydrocortisone.
Claim 16
Dependent Claim
23 words
The method of claim 12 wherein said colored protein is formed by mixing a protein and a protein dye in an aqueous solution.
Claim 17
Dependent Claim
18 words
The method of claim 14 wherein said protein has an average molecular weight of at least about 100,000.
Claim 18
Dependent Claim
24 words
The method of claim 14 wherein said dye is present in the range of about 0.0001 to 0.1% by weight of final assay mixture.
Claim 19
Dependent Claim
24 words
The method of claim 14 wherein said protein is present in an amount of at least 0.1% by weight of the final assay mixture.
Claim 20
Dependent Claim
24 words
The method of claim 12 wherein said precipitating agent is selected from the group consisting of polyethylene glycol, polylysine, dextran, polyvinylpyrolidone and polyvinyl alcohol.
Claim 21
Dependent Claim
22 words
The method of claim 12 wherein said precipitating agent is polyethylene glycol having an average molecular weight of at least about 6000.
Claim 22
Dependent Claim
26 words
The method of claim 12 wherein said precipitating agent is present in an amount of at least about 11/2% by weight of the final assay mixture.
Claim 23
Dependent Claim
14 words
The method of claim 12 wherein said labelled version of the antigen is radio-labelled.
Claim 24
Independent Claim
84 words
A method for performing an immunoassay, said method comprising incubating an aqueous mixture of (1) a sample containing an antigen to be assayed, (2) a known quantity of a labelled version of the antigen, (3) a prereacted first and second antibody for the antigen in an amount insufficient to react with all of the antigen, (4) a colored protein and (5) a precipitating agent that effects the precipitation of at least some of the colored protein, thus forming a colored precipitate for easy visualization.
Claim 25
Independent Claim
84 words
A method for performing an immunoassay, said method comprising incubating an aqueous mixture of (1) a sample containing an antigen to be assayed, (2) a known quantity of a labelled version of the antigen, (3) a prereacted first and second antibody for the antigen in an amount insufficient to react with all of the antigen, (4) a colored protein and (5) a precipitating agent that effects the precipitation of at least some of the colored protein, thus forming a colored precipitate for easy visualization.
Claim 26
Dependent Claim
21 words
The method of claim 22 wherein said antigen is selected from the group consisting of gentamicin, tobramycin, theophylline, digoxin, and hydrocortisone.
Claim 27
Dependent Claim
23 words
The method of claim 22 wherein said colored protein is formed by mixing a protein and a protein dye in an aqueous solution.
Claim 28
Dependent Claim
18 words
The method of claim 24 wherein said protein has an average molecular weight of at least about 100,000.
Claim 29
Dependent Claim
25 words
The method of claim 24 wherein said dye is present in the range of about 0.0001 to 0.1% by weight of the final assay mixture.
Claim 30
Dependent Claim
24 words
The method of claim 24 wherein said protein is present in an amount of at least 0.1% by weight of the final assay mixture.
Claim 31
Dependent Claim
24 words
The method of claim 22 wherein said precipitating agent is selected from the group consisting of polyethylene glycol, polylysine, dextran, polyvinylpyrolidone and polyvinyl alcohol.
Claim 32
Dependent Claim
22 words
The method of claim 22 wherein said precipitating agent is polyethylene glycol having an average molecular weight of at least about 6000.
Claim 33
Dependent Claim
26 words
The method of claim 22 wherein said precipitating agent is present in an amount of at least about 11/2% by weight of the final assay mixture.
Claim 34
Dependent Claim
14 words
The method of claim 22 wherein said labelled version of the antigen is radio-labelled.
Claim 35
Dependent Claim
97 words
A kit for performing the immunoassay for an antigen, said kit comprising, in combination: 1. a predetermined amount of a labelled version of the antigen to be assayed; 2. a predetermined amount of a first antibody to react with the antigen to be assayed to form a first antibody-antigen complex; 3. a predetermined amount of a second antibody to react with the first antibody-antigen complex to form a precipitate; 4. a predetermined amount of colored protein; and 5. a predetermined amount of a precipitating agent to effect the coprecipitation of at least some of the colored protein.
Claim 36
Dependent Claim
97 words
A kit for performing the immunoassay for an antigen, said kit comprising, in combination: 1. a predetermined amount of a labelled version of the antigen to be assayed; 2. a predetermined amount of a first antibody to react with the antigen to be assayed to form a first antibody-antigen complex; 3. a predetermined amount of a second antibody to react with the first antibody-antigen complex to form a precipitate; 4. a predetermined amount of colored protein; and 5. a predetermined amount of a precipitating agent to effect the coprecipitation of at least some of the colored protein.
Claim 37
Dependent Claim
22 words
The immunoassay kit of claim 32 wherein said antigen is selected from the group consisting of gentamicin, tobramycin, theophylline, digoxin, and hydrocortisone.
Claim 38
Dependent Claim
24 words
The immunoassay kit of claim 32 wherein said colored protein is formed by mixing a protein and a protein dye in an aqueous solution.
Claim 39
Dependent Claim
19 words
The immunoassay kit of claim 34 wherein said protein has an average molecular weight of at least about 100,000.
Claim 40
Dependent Claim
26 words
The immunoassay kit of claim 34 wherein said dye is present in the range of about 0.0001 to 0.1% by weight of the final assay mixture.
Claim 41
Dependent Claim
25 words
The immunoassay kit of claim 34 wherein said protein is present in an amount of at least 0.1% by weight of the final assay mixture.
Claim 42
Dependent Claim
25 words
The immunoassay kit of claim 32 wherein said precipitating agent is selected from the group consisting of polyethylene glycol, polylysine, dextran, polyvinylpyrolidone and polyvinyl alcohol.
Claim 43
Dependent Claim
23 words
The immunoassay kit of claim 32 wherein said precipitating agent is polyethylene glycol having an average molecular weight of at least about 6000.
Claim 44
Dependent Claim
27 words
The immunoassay kit of claim 32 wherein said precipitating agent is present in an amount of at least about 11/2% by weight of the final assay mixture.
Claim 45
Dependent Claim
15 words
The immunoassay kit of claim 32 wherein said labelled version of the antigen is radio-labelled.
Claim 46
Dependent Claim
70 words
A kit for performing the immunoassay for an antigen, said kit comprising, in combination: 1. a predetermined amount of a labelled version of the antigen to be assayed; 2. a predetermined amount of a prereacted first and second antibody for said antigen; 3. a predetermined amount of colored protein; and 4. a predetermined amount of a precipitating agent to effect the coprecipitation of at least some of the colored protein.
Claim 47
Dependent Claim
70 words
A kit for performing the immunoassay for an antigen, said kit comprising, in combination: 1. a predetermined amount of a labelled version of the antigen to be assayed; 2. a predetermined amount of a prereacted first and second antibody for said antigen; 3. a predetermined amount of colored protein; and 4. a predetermined amount of a precipitating agent to effect the coprecipitation of at least some of the colored protein.
Claim 48
Dependent Claim
22 words
The immunoassay kit of claim 42 wherein said antigen is selected from the group consisting of gentamicin, tobramycin, theophylline, digoxin, and hydrocortisone.
Claim 49
Dependent Claim
24 words
The immunoassay kit of claim 42 wherein said colored protein is formed by mixing a protein and a protein dye in an aqueous solution.
Claim 50
Dependent Claim
19 words
The immunoassay kit of claim 44 wherein said protein has an average molecular weight of at least about 100,000.
Claim 51
Dependent Claim
26 words
The immunoassay kit of claim 44 wherein said dye is present in the range of about 0.0001 to 0.1% by weight of the final assay mixture.
Claim 52
Dependent Claim
25 words
The immunoassay kit of claim 44 wherein said protein is present in an amount of at least 0.1% by weight of the final assay mixture.
Claim 53
Dependent Claim
25 words
The immunoassay kit of claim 42 wherein said precipitating agent is selected from the group consisting of polyethylene glycol, polylysine, dextran, polyvinylpyrolidone and polyvinyl alcohol.
Claim 54
Dependent Claim
23 words
The immunoassay kit of claim 42 wherein said precipitating agent is polyethylene glycol having an average molecular weight of at least about 6000.
Claim 55
Dependent Claim
27 words
The immunoassay kit of claim 42 wherein said precipitating agent is present in an amount of at least about 11/2% by weight of the final assay mixture.
Claim 56
Dependent Claim
15 words
The immunoassay kit of claim 42 wherein said labelled version of the antigen is radio-labelled.
Claim 57
Dependent Claim
38 words
The precipitate comprising in admixture a colored protein and a complex of an antigen, a first antibody for said antigen and a second antibody for said first antibody, wherein at least a portion of said antigen is labelled.
Claim 58
Dependent Claim
38 words
The precipitate comprising in admixture a colored protein and a complex of an antigen, a first antibody for said antigen and a second antibody for said first antibody, wherein at least a portion of said antigen is labelled.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-07-13 | 1985-01-12 | 1985-07-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-07-13 | 1989-01-12 | 1989-07-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-07-13 | 1993-01-12 | 1993-07-14 | $7700.00 | Window Closed / Lapsed |