US4427781A
Expired / Lapsed
US
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Particle agglutination immunoassay with agglutinator for determining haptens; PACIA
Assignee: de Duve Institute
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Filing: 1982-01-24
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Granted: 1984-01-24
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Expires: 2002-01-24
Abstract
Abstract A particle counting assay for haptens (small non-protein monovalent substances having a molecular weight below 1500) comprises mixing a liquid sample (e.g. of human origin) containing the hapten, with finely divided inert particles bearing the same hapten (or a specific analogue thereof), an agglutinator such as RF or C1q, and a measured amount of antibody, which amount is insufficient to cause agglutination of all the particles. The amount of hapten is determinable by measuring the extent of the agglutination.
Inventors
- 👤 Pierre L. Masson
- 👤 Daniel Collet-Cassart
- 👤 Carl G. Magnusson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 23:11
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/815; Y10S436/821; Y10S436/825
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
144 words
A method of assaying a hapten in a liquid sample, which comprises mixing with the liquid sample (i) finely divided inert particles bearing a reagent, (ii) an agglutinator and (iii) at least one other reagent, and determining the presence and/or amount of said hapten from the extent of agglutination of the particles, wherein: (a) the finely divided particles bear the same hapten as is under assay, or a specific analogue thereof; (b) said other reagent is an antibody against said hapten; (c) the amount of said antibody added to the mixture is less than the amount required to cause 100% agglutination of the particles; and (d) the agglutinator causes agglutination of hapten- or hapten analogue-bearing particles on which the antibody has formed a complex with said hapten or hapten analogue and does not cause agglutination of uncomplexed particles, uncomplexed hapten or free hapten-antibody complexes.
Claim 2
Independent Claim
144 words
A method of assaying a hapten in a liquid sample, which comprises mixing with the liquid sample (i) finely divided inert particles bearing a reagent, (ii) an agglutinator and (iii) at least one other reagent, and determining the presence and/or amount of said hapten from the extent of agglutination of the particles, wherein: (a) the finely divided particles bear the same hapten as is under assay, or a specific analogue thereof; (b) said other reagent is an antibody against said hapten; (c) the amount of said antibody added to the mixture is less than the amount required to cause 100% agglutination of the particles; and (d) the agglutinator causes agglutination of hapten- or hapten analogue-bearing particles on which the antibody has formed a complex with said hapten or hapten analogue and does not cause agglutination of uncomplexed particles, uncomplexed hapten or free hapten-antibody complexes.
Claim 3
Dependent Claim
26 words
A method according to claim 1, wherein the amount of antibody is such as to cause from 40 to 80% of the particles to become agglutinated.
Claim 4
Dependent Claim
18 words
A method according to claim 1, wherein the extent of agglutination is measured by counting the unagglutinated particles.
Claim 5
Dependent Claim
26 words
A method according to claim 1, wherein the hapten is selected from drugs, drug metabolites, intracellular messengers, hormones, vitamins, and haptens of bacteriological or virological origin.
Claim 6
Dependent Claim
17 words
A method according to claim 1, wherein the liquid sample is a human or animal body fluid.
Claim 7
Dependent Claim
17 words
A method according to claim 1, wherein the agglutinator is RF, C1lq, mouse serum or ascitic fluid.
Claim 8
Dependent Claim
52 words
A method according to claim 1, wherein the hapten under assay is a non-protein but wherein the liquid sample contains one or more proteins capable of interfering in the assay, wherein a protease enzyme is added to the liquid sample to digest said one or more proteins to remove their interfering effects.
Claim 9
Dependent Claim
30 words
A method according to claim 1, wherein one or more chaotropic agents are also added to said sample to reduce the effects of non-specific protein-protein interactions interfering in said assay.
Claim 10
Independent Claim
128 words
A method of assaying a human body fluid sample for a hapten selected from the group consisting of drugs, drug metabolites, intracellular messengers, hormones, vitamins, haptens of bacteriological origin and haptens of virological origin, which method comprises forming a mixture of (1) the liquid sample; (2) finely divided inert particles bearing the same hapten as is under assay or a specific analogue thereof; (3) an agglutinator selected from RF, C1q, ascitic fluid and mouse serum; and (4) a quantity of antibody against said hapten, which quantity causes from 40 to 80% of the said inert particles to agglutinate; incubating the mixture so formed; counting the number of particles remaining unagglutinated and therefrom determining the amount of hapten in the human body fluid sample by reference to standard results.
Claim 11
Independent Claim
128 words
A method of assaying a human body fluid sample for a hapten selected from the group consisting of drugs, drug metabolites, intracellular messengers, hormones, vitamins, haptens of bacteriological origin and haptens of virological origin, which method comprises forming a mixture of (1) the liquid sample; (2) finely divided inert particles bearing the same hapten as is under assay or a specific analogue thereof; (3) an agglutinator selected from RF, C1q, ascitic fluid and mouse serum; and (4) a quantity of antibody against said hapten, which quantity causes from 40 to 80% of the said inert particles to agglutinate; incubating the mixture so formed; counting the number of particles remaining unagglutinated and therefrom determining the amount of hapten in the human body fluid sample by reference to standard results.
Claim 12
Dependent Claim
12 words
A method according to claim 9, wherein the said hapten is digoxin.
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) | 1987-01-23 | 1987-07-25 | 1988-01-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-01-23 | 1991-07-25 | 1992-01-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-01-23 | 1995-07-25 | 1996-01-24 | $7700.00 | Window Closed / Lapsed |