US4581222A
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US
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Membrane immune assay
Assignee: California Institute of Technology
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Filing: 1983-02-09
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Granted: 1986-04-08
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Expires: 2003-02-09
Abstract
Abstract An immunoassay method for detection of antigen is disclosed. The method employs complement mediated lysis of vesicles loaded with In-111 or other gamma-emitting cation, and quantitative detection of the lysis by gamma-ray perturbed angular correlation (PAC) spectroscopy. The vesicles are labeled with a substance competitive to the antigen to be measured, and the concentration of the antigen in the sample measured by assessing the diminution in lysis due to the presence of the competing antigen. The method may also be used to assess the immunologic competence of a subject by injecting suitably sensitized vesicles and monitoring the in vivo lysis pattern by (PAC).
Inventors
- 👤 John D. Baldeschwieler
- 👤 Ronald C. Gamble
- 👤 Albert M. Lin
- 👤 George W. Tin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 11:36
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/534; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S436/827
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
64 words
A method for determining the concentration of an antigenic substance in a sample, which method comprises mixing together: (a) the sample; (b) antibody to said antigenic substance; (c) vesicles loaded with a gamma-emitting cation, and including on their surface an entity competitive with the antigenic substance for the antibody; and (d) complement; and measuring the time integrated pertubation factor associated with the gamma-emitting cation.
Claim 2
Independent Claim
64 words
A method for determining the concentration of an antigenic substance in a sample, which method comprises mixing together: (a) the sample; (b) antibody to said antigenic substance; (c) vesicles loaded with a gamma-emitting cation, and including on their surface an entity competitive with the antigenic substance for the antibody; and (d) complement; and measuring the time integrated pertubation factor associated with the gamma-emitting cation.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein the gamma-emitting cation is indium-111.
Claim 4
Dependent Claim
21 words
The method of claim 1 wherein the entity competitive with the antigen for antibody is the same entity as the antigen.
Claim 5
Dependent Claim
13 words
The method of claim 1 wherein the vesicles are large unilamellar vesicles (LUV).
Claim 6
Dependent Claim
18 words
The method of claim 1 wherein the antibody and complement are derived from the same species of organism.
Claim 7
Dependent Claim
24 words
The method of claim 1 wherein the mixture is incubated for between 15 minutes and 30 minutes before measuring the time integrated pertubation factor.
Claim 8
Dependent Claim
18 words
the method of claim 1 wherein the time integrated pertubation factor is measured as a function of time.
Claim 9
Dependent Claim
43 words
The method of claim 1 which includes a step comprising constructing a control by mixing loaded sensitized vesicles, antibody, and complement, determining the time integrated pertubation factor and comparing this time integrated perturbation factor to that of an analogous mixture which includes sample.
Claim 10
Independent Claim
46 words
A method for measuring the concentration of an antigenic substance in a sample, which method comprises: (a) mixing the sample with gamma-emitting cation loaded, antigen-sensitized vesicles, antibody to the antigenic substance, and complement; and (b) measuring the time integrated pertubation factor associated with the gamma-emitting cation.
Claim 11
Independent Claim
46 words
A method for measuring the concentration of an antigenic substance in a sample, which method comprises: (a) mixing the sample with gamma-emitting cation loaded, antigen-sensitized vesicles, antibody to the antigenic substance, and complement; and (b) measuring the time integrated pertubation factor associated with the gamma-emitting cation.
Claim 12
Dependent Claim
11 words
The method of claim 9 wherein the gamma-emitting cation is indium-111.
Claim 13
Dependent Claim
21 words
The method of claim 9 wherein the entity competitive with the antigen for antibody is the same entity as the antigen.
Claim 14
Dependent Claim
12 words
The method of claim 9 wherein the vesicles are large unilamellar vesicles.
Claim 15
Dependent Claim
18 words
The method of claim 9 wherein the antibody and complement are derived from the same species of organism.
Claim 16
Dependent Claim
24 words
The method of claim 9 wherein the mixture is incubated for between 15 minutes and 30 minutes before measuring the time integrated pertubation factor.
Claim 17
Dependent Claim
18 words
The method of claim 9 wherein the time integrated pertubation factor is measured as a function of time.
Claim 18
Dependent Claim
43 words
The method of claim 9 which includes a step comprising constructing a control by mixing loaded sensitized vesicles, antibody, and complement, determining the time integrated perturbation factor and comparing this time integrated pertubation factor to that of an analogous mixture which includes sample.
Claim 19
Independent Claim
44 words
A method for determining the immunological response in vivo in a subject vertebrate, which method comprises: (a) injecting a subject vertebrate with an antigen sensitized vesicles loaded with a gamma-emitting cation; and (b) measuring the time integrated pertubation factor for the gamma-emitting the cation.
Claim 20
Independent Claim
44 words
A method for determining the immunological response in vivo in a subject vertebrate, which method comprises: (a) injecting a subject vertebrate with an antigen sensitized vesicles loaded with a gamma-emitting cation; and (b) measuring the time integrated pertubation factor for the gamma-emitting the cation.
Claim 21
Dependent Claim
11 words
The method of claim 17 wherein the gamma-emitting cation is indium-111.
Claim 22
Dependent Claim
12 words
The method of claim 17 wherein the vesicles are large unilamellar vesicles.
Claim 23
Dependent Claim
18 words
The method of claim 17 wherein the time integrated pertubation factor is measured as a function of time.
References Cited (Backward Citations)
US4310506A
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1989-04-07 | 1989-10-07 | 1990-04-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-04-07 | 1993-10-07 | 1994-04-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-04-07 | 1997-10-07 | 1998-04-08 | $7700.00 | Window Closed / Lapsed |