US4388412A
Expired / Lapsed
US
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Immunoassay of phospholipid, such as phosphatidylcholine, in fluids such as amniotic
Assignee: HANA BIOLOGICS Inc A CORP OF
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Filing: 1982-02-02
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Granted: 1983-06-14
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Expires: 2002-02-02
Abstract
Abstract An immunologic assay for determining the presence of one or more phospholipids in a biological fluid. The method includes adding an ethanolic solution of diacylphosphatidylcholine or alkyltrimethylammonium halide and cholesterol to the biological fluid forming a macromolecular aggregate complex solution. To this solution is added antibody molecules to the phospholipids in an aqueous buffered medium causing an agglutination reaction. The product of the reaction is then examined to determine the presence of the phospholipids. The method is particularly useful in determining the presence of phosphatidylglycerol in a sample of amniotic fluid. Thus, the assay may be used in evaluating fetal lung maturity.
Inventors
- 👤 Kenichi K. Yabusaki
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 14:22
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/92; Y; Y10; Y10S; Y10S436/00; Y10S436/811; Y10S436/815
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
71 words
A method of determining the presence of a phospholipid in a biological fluid comprising: A. adding an ethanolic solution of diacylphosphatidylcholine and cholesterol to the biological fluid forming a macromolecular aggregate complex solution; B. adding the macromolecular aggregate complex solution to antibody molecules to the phospholipid in an aqueous buffered medium causing an agglutination reaction; and C. examining the results of the agglutination reaction to determine the presence of the phospholipid.
Claim 2
Independent Claim
71 words
A method of determining the presence of a phospholipid in a biological fluid comprising: A. adding an ethanolic solution of diacylphosphatidylcholine and cholesterol to the biological fluid forming a macromolecular aggregate complex solution; B. adding the macromolecular aggregate complex solution to antibody molecules to the phospholipid in an aqueous buffered medium causing an agglutination reaction; and C. examining the results of the agglutination reaction to determine the presence of the phospholipid.
Claim 3
Independent Claim
73 words
A method of determining the presence of a phospholipid in a biological fluid comprising: A. adding an ethanolic solution of an alkyltrimethylammonium halide and cholesterol to the biological fluid forming a macromolecular aggregate complex solution; B. adding the macromolecular aggregate complex solution to antibody molecules to the phospholipid in an aqueous buffered medium causing an agglutination reaction; and C. examining the results of the agglutination reaction to determine the presence of the phospholipid.
Claim 4
Independent Claim
73 words
A method of determining the presence of a phospholipid in a biological fluid comprising: A. adding an ethanolic solution of an alkyltrimethylammonium halide and cholesterol to the biological fluid forming a macromolecular aggregate complex solution; B. adding the macromolecular aggregate complex solution to antibody molecules to the phospholipid in an aqueous buffered medium causing an agglutination reaction; and C. examining the results of the agglutination reaction to determine the presence of the phospholipid.
Claim 5
Independent Claim
24 words
The method of claims 1 or 2 wherein the phospholipid is a member from the group consisting of phosphatidylcholine, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine and phosphatidylglycerol.
Claim 6
Independent Claim
14 words
The method of claims 1 or 2 wherein the biological fluid comprises amniotic fluid.
Claim 7
Dependent Claim
10 words
The method of claim 4 wherein the phospholipid comprises phosphatidylglycerol.
Claim 8
Dependent Claim
29 words
The method of claim 1 wherein sufficient diacylphosphatidylcholine and cholesterol are added to the biological fluid such that the phospholipid-containing macromolecular aggregates can be bound to the antibody molecules.
Claim 9
Dependent Claim
31 words
The method of claim 2 wherein sufficient alkyltrimethylammonium halide and cholesterol are added to the biological fluid such that the phospholipid containing macromolecular aggregates can be bound to the antibody molecules.
Claim 10
Independent Claim
97 words
A method of determining the presence of phosphatidylglycerol in a sample of amniotic fluid comprising: A. adding an ethanolic solution of diacylphosphatidylcholine and cholesterol to a sample of amniotic fluid; B. adding an additional quantity of amniotic fluid to the solution of subpart A forming macromolecular aggregates of lecithin, cholesterol and phosphatidylglycerol; C. forming an agglutination reaction by adding the phosphatidylglycerol macromolecular aggregate complex containing solution to a known quantity of antibody molecules to the phosphatidylglycerol in a buffered aqueous medium; and D. examining the results of the agglutination reaction to determine the presence of the phosphatidylglycerol.
Claim 11
Independent Claim
97 words
A method of determining the presence of phosphatidylglycerol in a sample of amniotic fluid comprising: A. adding an ethanolic solution of diacylphosphatidylcholine and cholesterol to a sample of amniotic fluid; B. adding an additional quantity of amniotic fluid to the solution of subpart A forming macromolecular aggregates of lecithin, cholesterol and phosphatidylglycerol; C. forming an agglutination reaction by adding the phosphatidylglycerol macromolecular aggregate complex containing solution to a known quantity of antibody molecules to the phosphatidylglycerol in a buffered aqueous medium; and D. examining the results of the agglutination reaction to determine the presence of the phosphatidylglycerol.
Claim 12
Independent Claim
99 words
A method of determining the presence of phosphatidylglycerol in a sample of amniotic fluid comprising: A. adding an ethanolic solution of alkyltrimethylammonium halide and cholesterol to a sample of amniotic fluid; B. adding an additional quantity of amniotic fluid to the solution of subpart A forming macromolecular aggregates of alkyltrimethylammonium halide, cholesterol and phosphatidylglycerol; C. forming an agglutination reaction by adding the phosphatidylglycerol macromolecular aggregate complex containing solution to a known quantity of antibody molecules to the phosphatidylglycerol in an aqueous buffered medium; and D. examining the results of the agglutination reaction to determine the presence of the phosphatidylglycerol.
Claim 13
Independent Claim
99 words
A method of determining the presence of phosphatidylglycerol in a sample of amniotic fluid comprising: A. adding an ethanolic solution of alkyltrimethylammonium halide and cholesterol to a sample of amniotic fluid; B. adding an additional quantity of amniotic fluid to the solution of subpart A forming macromolecular aggregates of alkyltrimethylammonium halide, cholesterol and phosphatidylglycerol; C. forming an agglutination reaction by adding the phosphatidylglycerol macromolecular aggregate complex containing solution to a known quantity of antibody molecules to the phosphatidylglycerol in an aqueous buffered medium; and D. examining the results of the agglutination reaction to determine the presence of the phosphatidylglycerol.
Claim 14
Independent Claim
28 words
The method of claims 8 or 9 wherein the buffered aqueous medium containing the antibodies to phosphatidylglycerol is buffered in the pH range of approximately 5 to 10.
Claim 15
Dependent Claim
28 words
The method of claim 8 or 9 wherein the buffered aqueous medium containing the antibodies to phosphatidylglycerol is buffered in the pH range of approximately 5.5 to 8.0.
Claim 16
Independent Claim
28 words
The method of claims 8 or 9 wherein the buffered aqueous medium containing the antibodies to phosphatidylglycerol is buffered in the pH range of approximately 6.5 to 7.0.
Claim 17
Independent Claim
17 words
The method of claims 8 or 9 wherein the aqueous medium is buffered with a phosphate buffer.
Claim 18
Independent Claim
15 words
The method of claims 1 or 2 wherein the complex solution further comprises sodium chloride.
Claim 19
Independent Claim
15 words
The method of claims 8 or 9 wherein the complex solution further comprises sodium chloride.
Claim 20
Dependent Claim
21 words
The method of claim 15 wherein the sodium chloride is present in an amount between approximately 0.01 to 5.0 weight percent.
Claim 21
Dependent Claim
21 words
The method of claim 15 wherein the sodium chloride is present in an amount between approximately 0.05 to 2.0 weight percent.
Claim 22
Dependent Claim
21 words
The method of claim 15 wherein the sodium chloride is present in an amount between approximately 0.5 to 1.0 weight percent.
Claim 23
Independent Claim
15 words
The method of claims 1 or 8 wherein the diacylphosphatidylcholine comprises hens egg yolk lecithin.
Claim 24
Independent Claim
14 words
The method of claims 2 or 9 wherein the alkyltrimethylammonium halide is hexadecyltrimethylammonium bromide.
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) | 1986-06-13 | 1986-12-13 | 1987-06-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-06-13 | 1990-12-13 | 1991-06-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-06-13 | 1994-12-13 | 1995-06-14 | $7700.00 | Window Closed / Lapsed |