US3887697A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Hemagglutination method for australia antigen and antibody thereto
Assignee: United States Department of Health and Human Services
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Filing: 1971-08-09
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Granted: 1975-06-03
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Expires: 1991-08-09
Abstract
Abstract Australia antigen or the antibody thereto is detected in the serum of a patient with viral hepatitis or detected in other appropriate biological material by a hemagglutination assay wherein indicator red cells are coated with purified hepatitisassociated-antigen and serum taken from the patient or other biological material is mixed therewith. If antibodies are present in the tested material hemagglutination will occur. In the test for the antigen the serum or other biological material is mixed with antibodies prior to contacting the mixture with the coated cells. Hemagglutination-inhibition indicates the presence of antigen in the test material.
Inventors
- 👤 Girish N Vyas
- 👤 Nahum Raphael Shulman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 14:38
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/576; Y; Y10; Y10S; Y10S436/00; Y10S436/82; Y10S436/826; Y10S530/00; Y10S530/806; Y10S530/81; Y10S530/827; Y10S530/829; Y10S530/83
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
104 words
A method for measuring Australia antigen in serum or other biological material comprising coating inert indicator human red blood cells with purified Australia antigen using chromic chloride as a coupling agent, suspending said coated cells in phosphate buffered saline solution containing bovine serum albumin, polyvinyl pyrrolidone, and polyoxyethylene sorbitan monooleate, adding an aliquot of a predetermined dilution of antibody to said antigen to each of serial two-fold dilutions of said serum or other material, and adding said suspension of coated cells to said serum-antibody mixture, whereby inhibition of hemagglutination in 1:2 dilution and higher dilutions indicates the presence of said antigen in said serum.
Claim 2
Independent Claim
104 words
A method for measuring Australia antigen in serum or other biological material comprising coating inert indicator human red blood cells with purified Australia antigen using chromic chloride as a coupling agent, suspending said coated cells in phosphate buffered saline solution containing bovine serum albumin, polyvinyl pyrrolidone, and polyoxyethylene sorbitan monooleate, adding an aliquot of a predetermined dilution of antibody to said antigen to each of serial two-fold dilutions of said serum or other material, and adding said suspension of coated cells to said serum-antibody mixture, whereby inhibition of hemagglutination in 1:2 dilution and higher dilutions indicates the presence of said antigen in said serum.
Claim 3
Independent Claim
105 words
A METHOD FOR MEASURING AUSTRALIA ANTIGEN IN A SERUM OR OTHER BIOLOGICAL MATERIAL COMPRISING COATING INERT INDICATOR HUMAN RED BLOOD CELLS WITH PURIFIED AUSTRALIA ANTIGEN USING CHROMIC CHLORIDE AS A COUPLING AGENT, SUSPENDING SAID COATED CELLS IN PHOSPHATE BUFFERED SALINE SOLUTION CONTAINING BOVINE SERUM ALBUMIN, POLYVINYL PYRROLIDONE, AND POLYOXYETHYLENE SORBITAN MONOOLEATE, ADDING AN ALIQUOT OF A PREDETERMINED DILUTION OF ANTIBODY TO SAID ANTIGEN TO EACH OF SERIAL TWO-FOLD DILUTIONS OF SAID SERUM OR OTHER MATERIAL, AND ADDING SAID SUSPENSION OF COATED CELLS TO SAID SERUM-ANTIBODY MIXTURE, WHEREBY INHIBITION OF HEMAGGLUTINATION IN 1:2 DILUTION AND HIGHER DILUTIONS INDICATES THE PRESENCE OF SAID ANTIGEN IN SAID SERUM.
Claim 4
Independent Claim
105 words
A METHOD FOR MEASURING AUSTRALIA ANTIGEN IN A SERUM OR OTHER BIOLOGICAL MATERIAL COMPRISING COATING INERT INDICATOR HUMAN RED BLOOD CELLS WITH PURIFIED AUSTRALIA ANTIGEN USING CHROMIC CHLORIDE AS A COUPLING AGENT, SUSPENDING SAID COATED CELLS IN PHOSPHATE BUFFERED SALINE SOLUTION CONTAINING BOVINE SERUM ALBUMIN, POLYVINYL PYRROLIDONE, AND POLYOXYETHYLENE SORBITAN MONOOLEATE, ADDING AN ALIQUOT OF A PREDETERMINED DILUTION OF ANTIBODY TO SAID ANTIGEN TO EACH OF SERIAL TWO-FOLD DILUTIONS OF SAID SERUM OR OTHER MATERIAL, AND ADDING SAID SUSPENSION OF COATED CELLS TO SAID SERUM-ANTIBODY MIXTURE, WHEREBY INHIBITION OF HEMAGGLUTINATION IN 1:2 DILUTION AND HIGHER DILUTIONS INDICATES THE PRESENCE OF SAID ANTIGEN IN SAID SERUM.
Claim 5
Dependent Claim
47 words
The method of claim 1, wherein said phosphate buffered saline solution is at a pH of 7.3, and contains 0.5 percent bovine serum albumin, 0.0025 percent polyvinyl pyrrolidone, and a 1:20,000 dilution of polyoxyethylene sorbitan monooleate in the form of an oily liquid of specific gravity 105-1.10.
Claim 6
Dependent Claim
85 words
The method of claim 1, wherein said red blood cells are coated with said purified antigen by suspending said cells as a 40 percent suspension in saline solution, mixing said cell suspension of cells in saline solution with three times the volume of purified antigen, adding an amount of 1.25 mM chromic chloride equal to the original volume of said cell suspension, agitating the resulting mixture for 5-20 minutes, washing the thus coated cells and suspending said coated cells in said phosphate buffered saline solution.
Claim 7
Dependent Claim
96 words
The method of claim 1, wherein said purified antigen is obtained by isolating antigen from plasma samples by combined rate and isopycnic zonal centrifugation with cesium chloride density gradients, and dialyzing against 0.85% saline solution at 4*C, said purified antigen having a Kjeldahl nitrogen content of 170 Mu g/ml and an optical density at 280 nm wave length of 0.445, said purified antigen further giving a single band in agarose at a maximum dilution of 1:2 with antiserum to the antigen and, when concentrated tenfold gives no precipitin band with rabbit antiserum against whole human plasma.
Claim 8
Independent Claim
107 words
A method for measuring antibody to Australia antigen in serum or other biological fluid comprising coating inert indicator human red blood cells with purified Australia antigen using chromic chloride as a coupling agent, suspending said coated cells in phosphate buffered saline solution containing bovine serum albumin, polyvinyl pyrrolidone, and polyoxyethylene sorbitan monooleate, and adding an aliquot of a predetermined dilution of said coated cells to each of serial two-fold dilutions of said serum or fluid in a microtiter plate, incubating the resulting mixtures, and centrifuging said microtiter plates whereby hemagglutination in serums with titers of at least 4 indicates the presence of said antibody in said serum.
Claim 9
Independent Claim
107 words
A method for measuring antibody to Australia antigen in serum or other biological fluid comprising coating inert indicator human red blood cells with purified Australia antigen using chromic chloride as a coupling agent, suspending said coated cells in phosphate buffered saline solution containing bovine serum albumin, polyvinyl pyrrolidone, and polyoxyethylene sorbitan monooleate, and adding an aliquot of a predetermined dilution of said coated cells to each of serial two-fold dilutions of said serum or fluid in a microtiter plate, incubating the resulting mixtures, and centrifuging said microtiter plates whereby hemagglutination in serums with titers of at least 4 indicates the presence of said antibody in said serum.
Claim 10
Dependent Claim
49 words
The method of claim 5, wherein said phosphate buffered saline solution is at a pH of 7.3, and contains 0.5 per cent bovine serum albumin, 0.0025 percent polyvinyl pyrrolidone, and a 1:20, 000 dilution of polyoxyethylene sorbitan monooleate in the Form of an oily liquid of specific gravity 1.05-1.10.
Claim 11
Dependent Claim
85 words
The method of claim 5, wherein said red blood cells are coated with said purified antigen by suspending said cells as a 40 percent suspension in saline solution, mixing said cell suspension of cells in saline solution with three times the volume of purified antigen, adding an amount of 1.25 mM chromic chloride equal to the original volume of said cell suspension, agitating the resulting mixture for 5-20 minutes, washing the thus coated cells and suspending said coated cells in said phosphate buffered saline solution.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-06-02 | 1978-12-02 | 1979-06-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-06-02 | 1982-12-02 | 1983-06-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-06-02 | 1986-12-02 | 1987-06-03 | $7700.00 | Window Closed / Lapsed |