US4320109A
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Immunoradiometric assay employing terminal radionuclide labeling and synthesis of conjugates for such assay
Assignee: University of Southern California
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Filing: 1979-06-29
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Granted: 1982-03-16
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Expires: 1999-06-29
Abstract
Abstract An immunoradiometric assay in which an antigen is reacted to its specific antibody, which complex is then separated and further reacted with a second, universal antibody. This second, universal antibody contains a chelating moiety, such as transferrin, capable of binding specifically and strongly short-lived radionuclides. The second complex, once formed is reacted with a radionuclide and the radio-activity is measured, determining thereby the quantity of the unknown antigen originally present.
Inventors
- 👤 Walter Wolf
- 👤 Robert M. Nakamura
- 👤 Ata Gokce
- 👤 Manuel Tubis
- 👤 Timothy J. O'Brien
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 19:44
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/534; G01N33/58; G01N33/60
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
108 words
In a method of immunoradiometric assay in which (a) an antigen and (b) an antibody specific to said antigen, are reacted to form a first complex which is reacted with a universal antibody for said complex from a species different from that of said specific antibody to form a second complex, and said second complex is separated from unreacted universal antibody and measured, the improvement according to which said universal antibody comprises a component capable of binding a radionuclide and including the steps of reacting a radionuclide with said separated second complex to bind said radionuclide to said universal antibody and measuring the radioactivity of said bound radionuclide.
Claim 2
Dependent Claim
108 words
In a method of immunoradiometric assay in which (a) an antigen and (b) an antibody specific to said antigen, are reacted to form a first complex which is reacted with a universal antibody for said complex from a species different from that of said specific antibody to form a second complex, and said second complex is separated from unreacted universal antibody and measured, the improvement according to which said universal antibody comprises a component capable of binding a radionuclide and including the steps of reacting a radionuclide with said separated second complex to bind said radionuclide to said universal antibody and measuring the radioactivity of said bound radionuclide.
Claim 3
Dependent Claim
21 words
The method of claim 1 wherein said universal antibody is of a species different from said antigen and said specific antibody.
Claim 4
Dependent Claim
12 words
The method of claim 1 wherein said component is a chelating agent.
Claim 5
Dependent Claim
18 words
The method of claim 3 wherein said chelating agent is of the group including transferrin, desferoxamine and D-penicillamine.
Claim 6
Independent Claim
19 words
The method of claims 2, 3 or 4 wherein said radionuclide is a radionuclide having a relatively short half-life.
Claim 7
Dependent Claim
13 words
The method of claim 2, 3 or 4 wherein said radionuclide is Indium-113.
Claim 8
Dependent Claim
10 words
The method of claim 1 wherein said component is transferrin.
Claim 9
Dependent Claim
16 words
The method of claim 7 wherein said radionuclide is a radionuclide having a relatively short half-life.
Claim 10
Dependent Claim
10 words
The method of claim 7 wherein said radionuclide is Indium-113.
Claim 11
Dependent Claim
71 words
In a method for complexing a radionuclide to a conjugate containing a component capable of binding a radionuclide comprising the steps of combining the chelate in a buffered solution with a salt of the radionuclide in the presence of nitriloacetic acid and a synergistic anion, the improvement comprising preparing said buffered solution using an eluting solvent for the radionuclide having an acid concentration sufficiently low to maintain said salt in solution.
Claim 12
Dependent Claim
71 words
In a method for complexing a radionuclide to a conjugate containing a component capable of binding a radionuclide comprising the steps of combining the chelate in a buffered solution with a salt of the radionuclide in the presence of nitriloacetic acid and a synergistic anion, the improvement comprising preparing said buffered solution using an eluting solvent for the radionuclide having an acid concentration sufficiently low to maintain said salt in solution.
Claim 13
Dependent Claim
12 words
The method of claim 10 wherein the component is a chelating agent.
Claim 14
Dependent Claim
18 words
The method of claim 11 wherein said chelating agent is of the group including transferrin, desferoxamine and D-penicillamine.
Claim 15
Dependent Claim
10 words
The method of claim 10 wherein the component is transferrin.
Claim 16
Independent Claim
19 words
The method of claims 10, 11 or 12 wherein said radionuclide is a radionuclide having a relatively short half-life.
Claim 17
Independent Claim
13 words
The method of claims 10, 11 or 12 wherein the radionuclide is Indium-113.
Claim 18
Independent Claim
211 words
A method of terminal labeling for radioimmunoassay comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first specie, specific to said antigen, and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second specie, specific to said antigen, and allowing said second antibody to react with said reacted amount of said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody from a specie different from said second specie, to said reacted amount of said second complex, said antibody being conjugated with a chelating moiety from the group including transferrin, desferoxamine and D-penicillamine said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding a radionuclide which will attach to said chelating moiety on said third complex and said third antibody; removing said excess unreacted amount of said third antibody; and measuring the radioactivity in said reacted amount of said third complex.
Claim 19
Independent Claim
211 words
A method of terminal labeling for radioimmunoassay comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first specie, specific to said antigen, and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second specie, specific to said antigen, and allowing said second antibody to react with said reacted amount of said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody from a specie different from said second specie, to said reacted amount of said second complex, said antibody being conjugated with a chelating moiety from the group including transferrin, desferoxamine and D-penicillamine said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding a radionuclide which will attach to said chelating moiety on said third complex and said third antibody; removing said excess unreacted amount of said third antibody; and measuring the radioactivity in said reacted amount of said third complex.
Claim 20
Dependent Claim
22 words
The method of claim 16 wherein said third antibody is of a species different from said second antibody and said first antibody.
Claim 21
Dependent Claim
11 words
The method of claim 16 wherein said radionuclide is short lived.
Claim 22
Dependent Claim
16 words
The method of claim 18 wherein said radionuclide has a half-life of less than 24 hours.
Claim 23
Independent Claim
216 words
A method of terminal labeling for radioimmunoassays comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first species, specific to said antigen and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second species, specific to said antigen, and allowing said second antibody to react with said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody, of a specie different from said second antibody, to said reacted amount of said second complex, said third antibody being conjugated with a chelating moiety from the group including transferrin, desferoxamine and D-penicillamine, said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding Indium-113m which will attach to said chelating moiety in said reacted amount of said third complex and said excess unreacted amount of said third antibody; removing said excess unreacted amount of said third antibody; and, measuring the radioactivity in said reacted amount of said third complex.
Claim 24
Independent Claim
216 words
A method of terminal labeling for radioimmunoassays comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first species, specific to said antigen and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second species, specific to said antigen, and allowing said second antibody to react with said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody, of a specie different from said second antibody, to said reacted amount of said second complex, said third antibody being conjugated with a chelating moiety from the group including transferrin, desferoxamine and D-penicillamine, said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding Indium-113m which will attach to said chelating moiety in said reacted amount of said third complex and said excess unreacted amount of said third antibody; removing said excess unreacted amount of said third antibody; and, measuring the radioactivity in said reacted amount of said third complex.
Claim 25
Independent Claim
210 words
A method of terminal labeling for radioimmunoassays comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first species, specific to said antigen and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second species, specific to said antigen, and allowing said second antibody to react with said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody, of a species different from said second antibody, to said reacted amount of said second complex, said third antibody being conjugated with a transferrin moiety, said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding Indium-113m which will attach to said transferrin moiety in said reacted amount of said third complex and said excess unreacted amount of said third antibody; removing said excess unreacted amount of said of said third antibody; and measuring the radioactivity in said reacted amount of said third complex.
Claim 26
Independent Claim
210 words
A method of terminal labeling for radioimmunoassays comprising the steps of: adding an unknown quantity of an antigen to an insoluble first antibody, from a first species, specific to said antigen and allowing all of said antigen to react with said first antibody to form a reacted amount of a first complex; adding a second antibody from a second species, specific to said antigen, and allowing said second antibody to react with said first complex to form a reacted amount of a second complex and an excess unreacted amount of said second antibody; removing said excess unreacted amount of said second antibody; adding a third antibody, of a species different from said second antibody, to said reacted amount of said second complex, said third antibody being conjugated with a transferrin moiety, said third antibody reacting with said reacted amount of said second complex to form a reacted amount of a third complex and an excess unreacted amount of said third antibody; adding Indium-113m which will attach to said transferrin moiety in said reacted amount of said third complex and said excess unreacted amount of said third antibody; removing said excess unreacted amount of said of said third antibody; and measuring the radioactivity in said reacted amount of said third complex.
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) | 1985-03-15 | 1985-09-14 | 1986-03-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-03-15 | 1989-09-14 | 1990-03-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-03-15 | 1993-09-14 | 1994-03-16 | $7700.00 | Window Closed / Lapsed |