US4591570A
Expired / Lapsed
US
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Matrix of antibody-coated spots for determination of antigens
Assignee: Tanox Inc
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Filing: 1983-02-02
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Granted: 1986-05-27
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Expires: 2003-02-02
Abstract
Abstract This invention relates to a novel immunoassay device and method for the determination of antigenic substances. The device essentially comprises a pattern or array of minute antibody-coated spots on the surface of a support. The array of antibody-coated spots is preferably in the form of a rectangular matrix. Each antibody-coated spot is made up of antibodies of a different and distinct specificity. A large number of different antibody-coated spots can be assembled on a very small portion of the surface of the support. The spots serve as tiny, specific immunoadsorbents of cells. The expression of particular surface antigen by cells may be detected by determining to which antibody-coated spot the cells bind.
Inventors
- 👤 Tse-Wen Chang
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 08:43
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54366; G01N33/544; G01N33/545; G01N33/551; G01N33/552; G01N33/569; G01N33/56966; G01N33/56977; B; B01; B01J; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00351; B01J2219/00364; B01J2219/00367; B01J2219/00583; B01J2219/00603; B01J2219/00605; B01J2219/0061; B01J2219/00612; B01J2219/00623; B01J2219/0063; B01J2219/00659; C; C40; C40B; C40B60/00; C40B60/14; Y; Y10; Y10S; Y10S435/00; Y10S435/965; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/809; Y10S436/815
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
100 words
An immunoassay device comprising a support having a substantially planar surface and having an array of small, discrete, closely spaced antibody-coated areas on the planar surface, the antibodies in the discrete, antibody-coated surface areas being absorbed to the planar surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated area containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated area and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete antibody-coated area.
Claim 2
Dependent Claim
100 words
An immunoassay device comprising a support having a substantially planar surface and having an array of small, discrete, closely spaced antibody-coated areas on the planar surface, the antibodies in the discrete, antibody-coated surface areas being absorbed to the planar surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated area containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated area and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete antibody-coated area.
Claim 3
Dependent Claim
21 words
An immunoassay device according to claim 1, wherein all of the discrete, antibody-coated areas have substantially the same size and shape.
Claim 4
Dependent Claim
24 words
An immunoassay device according to claim 1, wherein the antibodies in the discrete, antibody-coated areas are covalently bound to the surface of said support.
Claim 5
Dependent Claim
34 words
An immunoassay device according to claim 1, wherein the antibody in each of the discrete, antibody-coated surface areas, or in identifiable sets of said antibody-coated surface areas, is antibody of different and distinct specificity.
Claim 6
Dependent Claim
18 words
An immunoassay device according to claim 1, wherein the antibodies in the discrete, antibody-coated areas are monoclonal antibodies.
Claim 7
Dependent Claim
18 words
An immunoassay device according to claim 1, wherein the antibodies in the discrete, antibody-coated areas are monoclonal antibodies.
Claim 8
Dependent Claim
16 words
An immunoassay device according to claim 1, wherein the support is a glass or plastic coverslip.
Claim 9
Dependent Claim
18 words
An immunoassay device according to claim 1, wherein the array of discrete, antibody-coated areas is a rectangular matrix.
Claim 10
Dependent Claim
23 words
An immunoassay device according to claim 1, wherein discrete, antibody-coated surface areas are dots having diameters of about 0.25 mm to 1.0 mm.
Claim 11
Dependent Claim
29 words
An immunoassay device according to claim 1, wherein said antibody-coated dots on the surface of the support are arranged in a compact 5-20 dot by 5-20 dot rectangular matrix.
Claim 12
Dependent Claim
28 words
An immunoassay device according to claim 9, wherein the entire rectangular matrix is contained within an area of about 0.5-1.0 cm 2 of the surface of the support.
Claim 13
Independent Claim
155 words
A method of producing an immunoassay device which comprises the steps of: (a) applying droplets of substantially enriched aqueous solutions of antibodies of distinct specifications to a substantially planar surface of a support, locating said droplets so that they cover small, discrete, closely spaced areas of the planar surface of the support and form an array; (b) allowing said aqueous solutions of antibodies to remain on said areas of the surface for a period of time long enough for the antibodies in solution to be adsorbed to said areas of the surface to form discrete, antibody-coated areas; (c) swiftly flooding the entire surface of the solid support with an aqueous solution of protein in order to dilute the unadsorbed antibodies in the various solutions of antibodies and in order to block non-specific binding sites in areas of the surface of the solid support which surround the antibody-coated areas; and (d) washing and drying the device.
Claim 14
Independent Claim
155 words
A method of producing an immunoassay device which comprises the steps of: (a) applying droplets of substantially enriched aqueous solutions of antibodies of distinct specifications to a substantially planar surface of a support, locating said droplets so that they cover small, discrete, closely spaced areas of the planar surface of the support and form an array; (b) allowing said aqueous solutions of antibodies to remain on said areas of the surface for a period of time long enough for the antibodies in solution to be adsorbed to said areas of the surface to form discrete, antibody-coated areas; (c) swiftly flooding the entire surface of the solid support with an aqueous solution of protein in order to dilute the unadsorbed antibodies in the various solutions of antibodies and in order to block non-specific binding sites in areas of the surface of the solid support which surround the antibody-coated areas; and (d) washing and drying the device.
Claim 15
Dependent Claim
24 words
A method according to claim 12, wherein in step a the concentration of antibodies in said aqueous solutions of antibodies is about 10 μg/ml.
Claim 16
Dependent Claim
38 words
A method according to claim 12, wherein in step a each solution of antibodies is applied to the surface of the solid support in the form of a tiny droplet having a volume of about 0.005 μl-0.5 μl.
Claim 17
Dependent Claim
36 words
A method according to claim 12, wherein said small, discrete areas of the surface of the support to which said solutions of antibodies are applied are dots having diameters of about 0.25 mm to 1.0 mm.
Claim 18
Dependent Claim
23 words
A method according to claim 12, wherein said dots are located on the surface of the solid support to form a rectangular matrix.
Claim 19
Independent Claim
190 words
A method of determining the presence of particular antigens on the surface of cells, comprising the steps of: (a) applying a suspension of cells to a substantially planar surface of a support, said support having upon the planar surface an array of small, discrete, antibody-coated spots, each spot, or each of certain sets of spots, coated with antibodies of different and distinct specificity, the antibodies in the discrete, antibody-coated spots being adsorbed to the surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated spot containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated spot and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete, antibody-coated spot; (b) allowing the cells to settle upon the surface of the support so that the cells come into contact with all of the discrete antibody-coated spots; (c) allowing the cells to react with the antibodies in the discrete antibody-coated spots; (d) removing non-adherent cells; (e) determining to which discrete antibody-coated spots the cells have adhered.
Claim 20
Independent Claim
190 words
A method of determining the presence of particular antigens on the surface of cells, comprising the steps of: (a) applying a suspension of cells to a substantially planar surface of a support, said support having upon the planar surface an array of small, discrete, antibody-coated spots, each spot, or each of certain sets of spots, coated with antibodies of different and distinct specificity, the antibodies in the discrete, antibody-coated spots being adsorbed to the surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated spot containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated spot and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete, antibody-coated spot; (b) allowing the cells to settle upon the surface of the support so that the cells come into contact with all of the discrete antibody-coated spots; (c) allowing the cells to react with the antibodies in the discrete antibody-coated spots; (d) removing non-adherent cells; (e) determining to which discrete antibody-coated spots the cells have adhered.
Claim 21
Independent Claim
106 words
A method of determining the presence of particular antigens in a liquid comprising the steps of: (a) applying a sample of the liquid containing antigens to a substantially planar surface of a support, said support having upon its planar surface an array of small, discrete antibody-coated spots, each spot coated with antibodies of different and distinct specificity, so that the antigen containing liquid comes into contact with all of the discrete, antibody-coated spots; (b) allowing the antigens to react with the antibodies in the discrete, antibody-coated spots; (c) washing the device to remove all unbound antigens; (d) determining to which discrete, antibody-coated spots antigens are bound.
Claim 22
Independent Claim
106 words
A method of determining the presence of particular antigens in a liquid comprising the steps of: (a) applying a sample of the liquid containing antigens to a substantially planar surface of a support, said support having upon its planar surface an array of small, discrete antibody-coated spots, each spot coated with antibodies of different and distinct specificity, so that the antigen containing liquid comes into contact with all of the discrete, antibody-coated spots; (b) allowing the antigens to react with the antibodies in the discrete, antibody-coated spots; (c) washing the device to remove all unbound antigens; (d) determining to which discrete, antibody-coated spots antigens are bound.
Claim 23
Independent Claim
195 words
A method of determining the proportion of cells bearing specific surface differentiation antigens in a mixed population of cells comprising: (a) applying a suspension of cells to a substantially planar surface of a support, said support having upon its planar surface an array of small, discrete, antibody-coated spots, each spot coated with antibody of different and distinct specificity for cell surface differentiation antigens, the antibodies in the discrete, antibody-coated spots being adsorbed to the surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated spot containing antibodies which bind the antigen, the cells adhere tightly to the antibody-coated spot and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete, antibody-coated spot; (b) allowing the cells to settle upon the surface of the support so that the cells come into contact with all of the discrete, antibody-coated spots; (c) allowing the cells to react with the antibodies in the discrete, antibody-coated spots; (d) removing non-adherent cells; (e) quantitating cells bound to the discrete, antibody-coated spots; and (f) calculating the relative proportion of cells.
Claim 24
Independent Claim
195 words
A method of determining the proportion of cells bearing specific surface differentiation antigens in a mixed population of cells comprising: (a) applying a suspension of cells to a substantially planar surface of a support, said support having upon its planar surface an array of small, discrete, antibody-coated spots, each spot coated with antibody of different and distinct specificity for cell surface differentiation antigens, the antibodies in the discrete, antibody-coated spots being adsorbed to the surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated spot containing antibodies which bind the antigen, the cells adhere tightly to the antibody-coated spot and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete, antibody-coated spot; (b) allowing the cells to settle upon the surface of the support so that the cells come into contact with all of the discrete, antibody-coated spots; (c) allowing the cells to react with the antibodies in the discrete, antibody-coated spots; (d) removing non-adherent cells; (e) quantitating cells bound to the discrete, antibody-coated spots; and (f) calculating the relative proportion of cells.
Claim 25
Dependent Claim
37 words
An immunoassay device according to claim 1, for tissue typing, wherein the antibody in each of the discrete, antibody-coated areas, or in identifiable sets of the antibody-coated areas, is specific for a different and distinct transplantation antigen.
Claim 26
Dependent Claim
104 words
An immunoassay device for tissue typing comprising a solid support having a substantially planar surface and an array of small, discrete, closely-spaced anti-HLA antibody-coated spots on the planar surface, the anti-HLA antibody in the discrete, antibody-coated surface area being adsorbed to the planar surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated area containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated area and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete antibody-coated area.
Claim 27
Dependent Claim
104 words
An immunoassay device for tissue typing comprising a solid support having a substantially planar surface and an array of small, discrete, closely-spaced anti-HLA antibody-coated spots on the planar surface, the anti-HLA antibody in the discrete, antibody-coated surface area being adsorbed to the planar surface at a density and uniformity such that when cells bearing a surface antigen are brought into contact in sufficient concentration with a discrete, antibody-coated area containing antibodies which bind the antigen, the cells adhere tightly to the discrete, antibody-coated area and form a layer of bound cells which is substantially microscopically uniform over essentially all of the discrete antibody-coated area.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-05-26 | 1989-11-25 | 1990-05-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-05-26 | 1993-11-25 | 1994-05-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-05-26 | 1997-11-25 | 1998-05-27 | $7700.00 | Window Closed / Lapsed |