Patent US3843775A successfully refreshed from Google Patents.
US3843775A
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Radioimmunoassay of angiotensin and renin activity
Assignee: Mount Sinai Hospital Research Foundation Inc
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Filing: 1971-07-12
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Granted: 1974-10-22
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Expires: 1991-07-12
Abstract
Abstract A process for assaying the quantity of angiotensin and renin employing radioimmunoassay techniques, comprising the steps of combining solutions of (1) antibody to angiotensin coupled to poly-L-lysine, (2) angiotensin labeled with radioactivity, and (3) varying known amounts of unlabeled angiotensin to produce mixtures having a pH of about 7.0 to 9.5, which upon incubation form angiotensin-antibody complexes. The mixtures of angiotensinantibody complexes (antibody-bound angiotensin) and free angiotensin are applied either to paper strips, coated charcoal, ion exchange resins or resin sponges, and subjected to radioimmunoassay techniques to separate the antibody-bound angiotensin complexes from the free angiotensin. Some of the free and antibody-bound angiotensin is radioactive; and the ratio of radioactivity of the antibody-bound angiotensin complex to free angiotensin is measured and plotted as a function of the known concentrations of angiotensin in each mixture to prepare a standard concentration graph. The process is repeated employing solutions of a material to be assayed containing unknown amounts of unlabeled angiotensin, which may be derived from the action of renin on renin substrate, and the concentration of angiotensin is read from the graph.
Inventors
- 👤 R Wolf
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 20:08
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/74; G01N33/53; G01N33/536; G01N33/537; G01N33/539; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S436/815; Y10S436/817
Claims Analysis
Total Claims: 37
Claim 1
Independent Claim
13 words
A PROCESS FOR ASSAYING ANGIOTENSIN ACTIVITY IN A MATERIAL COMPRISING THE STEPS OF:
Claim 2
Dependent Claim
13 words
A PROCESS FOR ASSAYING ANGIOTENSIN ACTIVITY IN A MATERIAL COMPRISING THE STEPS OF:
Claim 3
Dependent Claim
25 words
PREPARING MIXTURES OF (I) ANTIBODY TO ANGIOTENSIN COUPLED TO POLY-L-LYSINE, (II) ANGIOTENSIN LABELED WITH A RADIOACTIVE ISOTOPE, AND (III) VARYING KNOWN AMOUNTS OF UNLABELED ANGIOTENSIN,
Claim 4
Dependent Claim
25 words
PREPARING MIXTURES OF (I) ANTIBODY TO ANGIOTENSIN COUPLED TO POLY-L-LYSINE, (II) ANGIOTENSIN LABELED WITH A RADIOACTIVE ISOTOPE, AND (III) VARYING KNOWN AMOUNTS OF UNLABELED ANGIOTENSIN,
Claim 5
Dependent Claim
26 words
INCUBATING THE MIXTURES TO FORM ANTIBODY-BOUND RADIOACTIVE ANGIOTENSIN COMPLEXES AND ANTIBODY-BOUND UNLABELED ANGIOTENSIN COMPLEXES AND INCREASES THE QUANTITY OF RADIOCATIVE FREE ANGIOTENSIN PRESENT IN THE MIXTURES,
Claim 6
Dependent Claim
26 words
INCUBATING THE MIXTURES TO FORM ANTIBODY-BOUND RADIOACTIVE ANGIOTENSIN COMPLEXES AND ANTIBODY-BOUND UNLABELED ANGIOTENSIN COMPLEXES AND INCREASES THE QUANTITY OF RADIOCATIVE FREE ANGIOTENSIN PRESENT IN THE MIXTURES,
Claim 7
Dependent Claim
26 words
incubating the mixtures to form antibody-bound radioactive angiotensin complexes and antibody-bound unlabeled angiotensin complexes and increase the quantity of radioactive free angiotensin present in the mixtures,
Claim 8
Dependent Claim
26 words
incubating the mixtures to form antibody-bound radioactive angiotensin complexes and antibody-bound unlabeled angiotensin complexes and increase the quantity of radioactive free angiotensin present in the mixtures,
Claim 9
Dependent Claim
38 words
A process according to claim 1, wherein the mixtures of (i) antibody, (ii) labeled angiotensin, and (iii) unlabeled angiotensin are prepared in a buffer solution and have a pH within the range of from about 7.0 to 9.5.
Claim 10
Dependent Claim
28 words
A process according to claim 1, wherein the mixtures of (i) antibody, (ii) labeled angiotensin, and (iii) unlabeled angiotensin are incubated at approximately 4*C for about 24 hours.
Claim 11
Dependent Claim
23 words
mixing the mixtures with a suspension of charcoal powder coated with dextran to cause the free angiotensin to react with the charcoal powder,
Claim 12
Dependent Claim
23 words
mixing the mixtures with a suspension of charcoal powder coated with dextran to cause the free angiotensin to react with the charcoal powder,
Claim 13
Dependent Claim
23 words
MIXING THE MIXTURES WITH A SUSPENSION OF CHARCOAL POWDER COATED WITH DEXTRAN TO CAUSE THE FREE ANGIOTENSIN TO REACT WITH THE CHARCOAL POWDER,
Claim 14
Dependent Claim
23 words
MIXING THE MIXTURES WITH A SUSPENSION OF CHARCOAL POWDER COATED WITH DEXTRAN TO CAUSE THE FREE ANGIOTENSIN TO REACT WITH THE CHARCOAL POWDER,
Claim 15
Dependent Claim
17 words
PRECIPITATING THE CHARCOAL POWDER FROM THE SUSPENSION TO SEPARATE THE FREE ANGIOTENSIN FROM THE ANTIBODY-BOUND ANGIOTENSIN COMPLEXES,
Claim 16
Dependent Claim
17 words
PRECIPITATING THE CHARCOAL POWDER FROM THE SUSPENSION TO SEPARATE THE FREE ANGIOTENSIN FROM THE ANTIBODY-BOUND ANGIOTENSIN COMPLEXES,
Claim 17
Dependent Claim
17 words
precipitating the charcoal powder from the suspension to separate the free angiotensin from the antibody-bound angiotensin complexes,
Claim 18
Dependent Claim
17 words
precipitating the charcoal powder from the suspension to separate the free angiotensin from the antibody-bound angiotensin complexes,
Claim 19
Dependent Claim
43 words
A process according to claim 1, wherein mixtures of (i) antibody and (ii) labeled angiotensin are prepared and diluted to yield an initial bound to free angiotensin radioactivity ratio of from approximately 1.5 to 3.0 prior to being added to the unlabeled angiotensin.
Claim 20
Dependent Claim
14 words
A process according to claim 1, wherein the charcoal powder is precipitated by centrifugation.
Claim 21
Dependent Claim
29 words
counting the radioactivity of the supernatant fluid and of the charcoal powder residue to determine the ratios of the radioactivity of antibody-bound angiotensin to the radioactivity of free angiotensin,
Claim 22
Dependent Claim
29 words
counting the radioactivity of the supernatant fluid and of the charcoal powder residue to determine the ratios of the radioactivity of antibody-bound angiotensin to the radioactivity of free angiotensin,
Claim 23
Dependent Claim
29 words
COUNTING THE RADIOACTIVITY OF THE SUPERNATURAL FLUID AND OF THE CHARCOAL POWDER RESIDUE TO DETERMINE THE RATIOS OF THE RADIOACTIVITY OF ANTIBODY-BOUND ANGIOTENSIN TO THE RADIOACTIVITY OF FREE ANGIOTENSIN,
Claim 24
Dependent Claim
29 words
COUNTING THE RADIOACTIVITY OF THE SUPERNATURAL FLUID AND OF THE CHARCOAL POWDER RESIDUE TO DETERMINE THE RATIOS OF THE RADIOACTIVITY OF ANTIBODY-BOUND ANGIOTENSIN TO THE RADIOACTIVITY OF FREE ANGIOTENSIN,
Claim 25
Dependent Claim
35 words
REFERENCING THE RATIOS OF BOUND TO FREE RADIOACTIVE ANGIOTENSIN TO THE KNOWN AMOUNTS OF ANGIOSTENSIN IN SAID MIXTURES TO ESTABLISH A RELATIONSHIP BETWEEN THE RATIO OF BOUND TO FREE ANGIOTENSIN AND THE CONCENTRATION OF ANGIOTENSIN,
Claim 26
Dependent Claim
35 words
REFERENCING THE RATIOS OF BOUND TO FREE RADIOACTIVE ANGIOTENSIN TO THE KNOWN AMOUNTS OF ANGIOSTENSIN IN SAID MIXTURES TO ESTABLISH A RELATIONSHIP BETWEEN THE RATIO OF BOUND TO FREE ANGIOTENSIN AND THE CONCENTRATION OF ANGIOTENSIN,
Claim 27
Dependent Claim
35 words
referencing the ratios of bound to free radioactive angiotensin to the known amounts of angiotensin in said mixtures to establish a relationship between the ratio of bound to free angiotensin and the concentration of angiotensin,
Claim 28
Dependent Claim
35 words
referencing the ratios of bound to free radioactive angiotensin to the known amounts of angiotensin in said mixtures to establish a relationship between the ratio of bound to free angiotensin and the concentration of angiotensin,
Claim 29
Dependent Claim
122 words
A process according to claim 1 further comprising the steps of: incubating a duplicate quantity of the solution of material to be assayed for approximately 3 hours at about 37*C prior to adding thereto the labeled angiotensin and angiotensin antibody, ascertaining the concentration of angiotensin in the duplicate solution by repeating steps (1) to (5) using the incubated duplicate solution in place of the known amounts of unlabeled angiotensin to determine the ratio of radioactive bound angiotensin to radioactive free angiotensin for the duplicate solution, and locating the ratio thus determined on the line graph plotted according to step (6), whereby an indication of renin activity in the material may be obtained from the rate of release of angiotensin by the material.
Claim 30
Dependent Claim
46 words
repeating steps (1) to (5) using a solution of the material to be assayed containing an unknown amount of unlabeled angiotensin in place of the known amounts of unlabeled angiotensin to determine the ratio of radioactive bound angiotensin to radioactive free angiotensin for the material, and
Claim 31
Dependent Claim
46 words
repeating steps (1) to (5) using a solution of the material to be assayed containing an unknown amount of unlabeled angiotensin in place of the known amounts of unlabeled angiotensin to determine the ratio of radioactive bound angiotensin to radioactive free angiotensin for the material, and
Claim 32
Dependent Claim
47 words
REPEATING STEPS (1) TO (5) USING A SOLUTION OF THE MATERIAL TO BE ASSAYED CONTAINING AN UNKNOWN AMOUNT OF UNLABELED ANGIOTENSIN IN PLACE OF THE KNOWN AMOUNTS OF UNLA BELED ANGIOTENSIN TO DETERMINE THE RATIO OF RADIOACTIVE BOUND ANGIOTENSIN TO RADIOACITVE FREE ANGIOTENSIN FOR THE MATERIAL, AND
Claim 33
Dependent Claim
47 words
REPEATING STEPS (1) TO (5) USING A SOLUTION OF THE MATERIAL TO BE ASSAYED CONTAINING AN UNKNOWN AMOUNT OF UNLABELED ANGIOTENSIN IN PLACE OF THE KNOWN AMOUNTS OF UNLA BELED ANGIOTENSIN TO DETERMINE THE RATIO OF RADIOACTIVE BOUND ANGIOTENSIN TO RADIOACITVE FREE ANGIOTENSIN FOR THE MATERIAL, AND
Claim 34
Dependent Claim
28 words
COMPARING THE RATIO DETERMINED ACCORDING TO STEP (7) WITH THE RELATIONSHIP PREVIOUSLY ESTABLISHED ACCORDING TO STEP (6) TO DETERMINE THE CONCENTRATION OF ANGIOTENSIN IN THE MATERIAL BEING ASSAYED.
Claim 35
Dependent Claim
28 words
COMPARING THE RATIO DETERMINED ACCORDING TO STEP (7) WITH THE RELATIONSHIP PREVIOUSLY ESTABLISHED ACCORDING TO STEP (6) TO DETERMINE THE CONCENTRATION OF ANGIOTENSIN IN THE MATERIAL BEING ASSAYED.
Claim 36
Dependent Claim
28 words
comparing the ratio determined according to step (7) with the relationship previously established according to step (6) to determine the concentration of angiotensin in the material being assayed.
Claim 37
Dependent Claim
28 words
comparing the ratio determined according to step (7) with the relationship previously established according to step (6) to determine the concentration of angiotensin in the material being assayed.
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) | 1977-10-21 | 1978-04-22 | 1978-10-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1981-10-21 | 1982-04-22 | 1982-10-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1985-10-21 | 1986-04-22 | 1986-10-22 | $7700.00 | Window Closed / Lapsed |