Patent US4469787A successfully refreshed from Google Patents.
US4469787A
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US
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Immunoassay involving soluble complex of second antibody and labeled binding protein
Assignee: BYK-Sangtec Diagnostica GmbH and Co KG
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Filing: 1982-05-14
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Granted: 1984-09-04
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Expires: 2002-05-14
Abstract
Abstract A method and kit for determining the presence of a polyvalent ligand in an aqueous fluid. The method involves incubating the fluid with an immobilized antibody to the ligand to form an immobilized ligand-antibody complex, then incubating the immobilized ligand-antibody complex with a solution of a soluble complex of a second antibody (to the ligand) and a labeled binding protein, such as protein A. The labeled binding protein is specifically bound to the Fc portion of the second antibody. The unbound second antibody and labeled binding protein are washed from the immobilized complex, and the presence of bound labeled binding protein is determined as a measure of the concentration of the ligand in the aqueous fluid.
Inventors
- 👤 James W. Woods
- 👤 John N. St. Denis
- 👤 Dana A. Chapman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 12:05
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6854; G01N33/6857; G01N33/53; G01N33/543; G01N33/54306; G01N33/563; G01N33/573; G01N33/74; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/975; Y10S436/00; Y10S436/801; Y10S436/804; Y10S436/807; Y10S436/808; Y10S436/811; Y10S436/813; Y10S436/815; Y10S436/817; Y10S436/818; Y10S436/828
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
173 words
A method for quantitatively determining the presence of polyvalent ligand in an aqueous fluid which comprises: (a) incubating said fluid with a solid support having a first antibody to said ligand bound thereto to form an immobilized antibody-ligand complex; (b) incubating said immobilized antibody-ligand complex with a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein specifically bound to the F c portion of said second antibody; (c) washing unbound antibody and labeled binding protein from the solid support; and (d) determining the presence of labeled binding protein bound to said solid support or remaining in solution, as a measure of the concentration of the ligand in the aqueous fluid; wherein said method is further characterized in that the solution of second antibody-labeled binding protein complex contains an excess concentration of second antibody to provide a substantially linear relationship between the concentration of ligand and the amount of labeled binding protein bound to the solid support, while maintaining analytically acceptable low levels of background interference.
Claim 2
Independent Claim
173 words
A method for quantitatively determining the presence of polyvalent ligand in an aqueous fluid which comprises: (a) incubating said fluid with a solid support having a first antibody to said ligand bound thereto to form an immobilized antibody-ligand complex; (b) incubating said immobilized antibody-ligand complex with a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein specifically bound to the F c portion of said second antibody; (c) washing unbound antibody and labeled binding protein from the solid support; and (d) determining the presence of labeled binding protein bound to said solid support or remaining in solution, as a measure of the concentration of the ligand in the aqueous fluid; wherein said method is further characterized in that the solution of second antibody-labeled binding protein complex contains an excess concentration of second antibody to provide a substantially linear relationship between the concentration of ligand and the amount of labeled binding protein bound to the solid support, while maintaining analytically acceptable low levels of background interference.
Claim 3
Dependent Claim
24 words
The method of claim 1, wherein the molar ratio of soluble second antibody to labeled binding protein is from about 250:1 to about 5,000:1.
Claim 4
Dependent Claim
24 words
The method of claim 1, wherein the molar ratio of soluble second antibody to labeled binding protein is from about 50:1 to about 24,000:1.
Claim 5
Dependent Claim
17 words
The method of claim 3, wherein the labeled binding protein is protein A obtained from Staphylococcus aureus.
Claim 6
Dependent Claim
10 words
The method of claim 4, wherein the ligand is ferritin.
Claim 7
Dependent Claim
34 words
The method of claim 4, wherein the ligand is selected from the group consisting of ferritin, thyroid stimulating hormone, creatinine phosphokinase, prostatic acid-phosphatase, and human chorionic gonadotroin, alpha fetoprotein, parathyroid hormone, insulin and C-peptide.
Claim 8
Dependent Claim
13 words
The method of claim 6, wherein the first antibody is an intact antibody.
Claim 9
Dependent Claim
29 words
The method of claim 5 or 6, wherein the labeled binding protein is selected from the group consisting of 125 I-Protein A, 3 H-Protein A and 14 C-Protein A.
Claim 10
Dependent Claim
16 words
The method of claim 5 or 6, wherein the labeled binding protein is enzyme-labeled Protein A.
Claim 11
Dependent Claim
20 words
The method of claim 5 or 6, wherein the labeled binding protein is protein A labeled with a fluorescent substance.
Claim 12
Dependent Claim
75 words
An immunoassay test kit for determining the presence of a ligand in an aqueous fluid comprising a solid support coated with an antibody to said ligand and, in a separate container, a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein, wherein the concentrations of second antibody and labeled binding protein in said solution are in a molar ratio of from about 50:1 to about 24,000:1 respectively.
Claim 13
Dependent Claim
75 words
An immunoassay test kit for determining the presence of a ligand in an aqueous fluid comprising a solid support coated with an antibody to said ligand and, in a separate container, a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein, wherein the concentrations of second antibody and labeled binding protein in said solution are in a molar ratio of from about 50:1 to about 24,000:1 respectively.
Claim 14
Dependent Claim
24 words
The test kit of claim 11, wherein the molar ratio of second antibody to labeled binding protein is from about 250:1 to about 5,000:1.
Claim 15
Dependent Claim
15 words
The test kit of claim 12, wherein the labeled binding protein is radiolabeled Protein A.
Claim 16
Dependent Claim
30 words
The test kit of claim 12, wherein the labeled binding protein is enzyme-labeled Protein A, and said test kit further comprises a solution of a chromogenic substrate for said enzyme.
Claim 17
Independent Claim
141 words
A method for quantitatively determining the presence of polyvalent ligand in an aqueous fluid which comprises: (a) incubating said fluid with a solid support having a first antibody to said ligand bound therto to form an immobilized antibody-ligand complex; (b) incubating said immobilized antibody-ligand complex with a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein specifically bound to the Fc portion of said second antibody; (c) washing unbound antibody and labeled binding protein from the solid support; and (d) determining the presence of labeled binding protein bound to said solid support or remaining in solution, as a measure of the concentration of the ligand in the aqueous fluid; wherein said method is characterized in that the first antibody is an IgG immunoglobin which has been enzymatically digested to remove the Fc portion.
Claim 18
Independent Claim
141 words
A method for quantitatively determining the presence of polyvalent ligand in an aqueous fluid which comprises: (a) incubating said fluid with a solid support having a first antibody to said ligand bound therto to form an immobilized antibody-ligand complex; (b) incubating said immobilized antibody-ligand complex with a solution containing a soluble complex of a second antibody to said ligand and a labeled binding protein specifically bound to the Fc portion of said second antibody; (c) washing unbound antibody and labeled binding protein from the solid support; and (d) determining the presence of labeled binding protein bound to said solid support or remaining in solution, as a measure of the concentration of the ligand in the aqueous fluid; wherein said method is characterized in that the first antibody is an IgG immunoglobin which has been enzymatically digested to remove the Fc portion.
Claim 19
Dependent Claim
24 words
The method of claim 15, wherein the molar ratio of soluble second antibody to labeled binding protein is from about 500:1 to about 1:1.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1987-09-04 | 1988-03-05 | 1988-09-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-09-04 | 1992-03-05 | 1992-09-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-09-04 | 1996-03-05 | 1996-09-04 | $7700.00 | Window Closed / Lapsed |