Patent US4853335A successfully refreshed from Google Patents.
US4853335A
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Colloidal gold particle concentration immunoassay
Assignee: Aberlyn Capital Management LP
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Filing: 1987-09-28
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Granted: 1989-08-01
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Expires: 2007-09-28
Abstract
Abstract A sandwich immunoassay method is disclosed that is useful for the identification of an antigen in biological specimens. The method is comprised of a colloidal gold labeled binder specific for epitopes or receptors of a ligand, nonporous particulate solid phase which have antiligand covalently attached, and which said reagents are combined with the biological specimen or an extract of the biological specimen, and after an appropriate incubation, the said reactant mixture is contacted to a porous film having an exposed surface area of contact no greater than 30 mm 2 to separate and concentrate the particle bound colloidal gold labeled binder from the unbound. The identification of an antigen is determined by the presence of color on the surface of the solid phase particles captured on the surface of the film. A competitive inhibition assay can be also used to identify the presence of a hapten or antigen, in which case the presence of the analyte is determined by the absence of color.
Inventors
- 👤 Duane A. Olsen
- 👤 David Bernstein
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 18:12
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/585; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S436/00; Y10S436/807
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
134 words
A method for detecting in a biological specimen the presence of a ligand or an antiligand selected from the group consisting of antigens, haptens, antibodies and nucleic acids, said method comprising the steps of: (a) adding to said biological specimen (1) a colloidal gold labeled ligand or a colloidal gold labeled antiligand and (2) solid phase capture particles coated with ligand or with antiligand; (b) applying the resultant of step (a) to a small zone on a porous film, said film having a minimal effective pore size smaller than (2) but greater than the size of (1), whereby the capture particles (2) are deposited on the exposed surface of the porous film; and (c) observing the small zone of the porous film for any color development due to (1) having become bound to (2).
Claim 2
Independent Claim
134 words
A method for detecting in a biological specimen the presence of a ligand or an antiligand selected from the group consisting of antigens, haptens, antibodies and nucleic acids, said method comprising the steps of: (a) adding to said biological specimen (1) a colloidal gold labeled ligand or a colloidal gold labeled antiligand and (2) solid phase capture particles coated with ligand or with antiligand; (b) applying the resultant of step (a) to a small zone on a porous film, said film having a minimal effective pore size smaller than (2) but greater than the size of (1), whereby the capture particles (2) are deposited on the exposed surface of the porous film; and (c) observing the small zone of the porous film for any color development due to (1) having become bound to (2).
Claim 3
Dependent Claim
23 words
The method of claim 1 carried in a non-competitive mode wherein a positive result is indicated by color development on the porous film.
Claim 4
Dependent Claim
27 words
The method of claim 1 carried out in a competitive mode wherein a positive result is indicated by an absence of color development on the porous film.
Claim 5
Dependent Claim
28 words
The method of claim 1, wherein a substep of adding solvent or a buffer to said capture particles after performing step (b) and prior to performing step (c).
Claim 6
Dependent Claim
21 words
The method of claim 1, wherein said capture particles have a mean diameter of from about 0.10 microns to 20.0 microns.
Claim 7
Dependent Claim
26 words
The method of claim 1, wherein said small zone on porous film having an area of from about 0.2 mm 2 to 30 mm 2 .
Claim 8
Dependent Claim
22 words
The method of claim 1, wherein said colloidal gold having a mean particle diameter of from about 5 nm to 10 nm.
Claim 9
Dependent Claim
38 words
The method of claim 1, wherein the fluid phase of said resultant step (a) flows into said porous film by a process selected from the group comprised of diffusion, positive pressure, negative pressure, filtration or any combination thereof.
Claim 10
Dependent Claim
17 words
The method of claim 1, wherein the ligands or antiligands are covalently attached to said capture particles.
Claim 11
Dependent Claim
28 words
The method of claim 1, wherein the antiligand bound to said capture particles is selected from the group comprised of antibodies, lectins, receptors, DNA fragments, and RNA fragments.
Claim 12
Dependent Claim
24 words
The method of claim 1, wherein the said antiligand is a DNA or RNA fragment and the said ligand is complimentary DNA or RNA.
Claim 13
Dependent Claim
36 words
The method of claim 1, wherein step (a) has an additional substep of treating said biological speciment with a solubilizing agent selected from the group comprised of enzymes, detergents, salts, reducing reagents, and any combination thereof.
Claim 14
Dependent Claim
17 words
The method of claim 1, wherein (1) and (2) are mixed and added as a single reagent.
Claim 15
Dependent Claim
25 words
The method of claim 1, wherein the resultant of step (a) is incubated from about 1 second to 18 hours prior to performing step (b).
Claim 16
Independent Claim
138 words
A method for detecting in a biological specimen the presence of a ligand or an antiligand selected from the group consisting of antigens, haptens, antibodies and nucleic acids, said method comprising the steps of: (a) applying (2) solid phase capture particles coated with ligands or with antiligands to a small zone or a porous film, said film having a minimal effective pore size smaller than said capture particles, (b) adding to said biological specimen (1) a colloidal gold labeled ligand or a colloidal gold labeled antiligand, said colloidal gold having a mean particle size smaller than said minimal effective pore size of said film, and (c) applying the resultant of (b) to the resultant of (a) and (d) observing the small zone of the porous film for any color development due to (1) having become bound to (2).
Claim 17
Independent Claim
138 words
A method for detecting in a biological specimen the presence of a ligand or an antiligand selected from the group consisting of antigens, haptens, antibodies and nucleic acids, said method comprising the steps of: (a) applying (2) solid phase capture particles coated with ligands or with antiligands to a small zone or a porous film, said film having a minimal effective pore size smaller than said capture particles, (b) adding to said biological specimen (1) a colloidal gold labeled ligand or a colloidal gold labeled antiligand, said colloidal gold having a mean particle size smaller than said minimal effective pore size of said film, and (c) applying the resultant of (b) to the resultant of (a) and (d) observing the small zone of the porous film for any color development due to (1) having become bound to (2).
Claim 18
Dependent Claim
23 words
The method of claim 15 carried in a non-competitive mode wherein a positive result is indicated by color development on the porous film.
Claim 19
Dependent Claim
27 words
The method of claim 15 carried out in a competitive mode wherein a positive result is indicated by an absence of color development on the porous film.
Claim 20
Dependent Claim
25 words
The method of claim 15, wherein a substep of adding solvent or buffer to said capture particles after step (c) and prior to step (d).
Claim 21
Dependent Claim
21 words
The method of claim 15, wherein said capture particles having a mean diameter of from about 0.10 microns to 20.0 microns.
Claim 22
Dependent Claim
26 words
The method of claim 15, wherein said small zone on porous film having an area of from about 0.2 mm 2 to 30 mm 2 .
Claim 23
Dependent Claim
22 words
The method of claim 15, wherein said colloidal gold having a mean particle diameter of from about 5 nm to 100 nm.
Claim 24
Dependent Claim
38 words
The method of claim 15, wherein the fluid phase of said resultant step (a) flows into said porous film by a process selected from the group comprised of diffusion, positive pressure, negative pressure, filtration or any combination thereof.
Claim 25
Dependent Claim
17 words
The method of claim 15, wherein the ligands or antiligands are covalently attached to said capture particles.
Claim 26
Dependent Claim
28 words
The method of claim 15, wherein the antiligand bound to said capture particles is selected from the group comprised of antibodies, lectins, receptors, DNA fragments, and RNA fragments.
Claim 27
Dependent Claim
24 words
The method of claim 15, wherein the said antiligand is a DNA or RNA fragment and the said ligand is complimentary DNA or RNA.
Claim 28
Dependent Claim
36 words
The method of claim 15, wherein step (a) has an additional substep of treating said biological specimen with a solubilizing agent selected from the group comprised of enzymes, detergents, salts, reducing reagents, and any combination thereof.
Claim 29
Dependent Claim
25 words
The method of claim 15, wherein the resultant of step (b) is incubated from about 1 second to 18 hours prior to performing step (c).
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