Patent US4108975A successfully refreshed from Google Patents.
US4108975A
Expired / Lapsed
US
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Radioimmunoassay system
Assignee: Becton Dickinson and Co
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Filing: 1977-03-04
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Granted: 1978-08-22
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Expires: 1997-03-04
Abstract
Abstract An improved radioimmunoassey procedure involves the use of an improved reusable immunoadsorbent through which a sample of an antigen and radioactive labelled antigen is flowed to form a bound and unbound fraction. The bound fraction is releasably coupled to the immunoadsorbent which includes a solid particulate substrate to which antibodies, specific to the antigen, are covalently bonded by reaction between the carboxyl or amine group of the antibody and a group such as an acyl azide, carbonate, thiocarbonate, polythiol, isocyanate, epoxide and chlorothioformate group, for example, reacted on the substrate. The substrate includes a porous refractory material having at least one polymeric material bonded thereto as a barrier and to which is linked one of the above groups for reaction with the carboxyl or amine group of the antibody. The antigen bound to the immunoadsorbent is stoichiometrically released by an eluting solvent and the immunoadsorbent is reused. Concentration of the antigen is determined with reference to one of the bound or unbound fractions, or both.
Inventors
- 👤 Richard Harold Hales
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 14:47
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54306; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S436/807; Y10S436/808; Y10S436/815; Y10S436/817
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
236 words
In a method of radioimmunoassay wherein an antigen bearing sample and a radioactive labelled antigen are brought into contact with an immobilized immunoadsorbent to form a fraction bound to the immunoadsorbent and an unbound fraction and wherein the concentration of the antigen is determined as a function of the bound or unbound fraction, or both, the improvement comprising: flowing an antigen bearing sample and a radioactive labelled antigen into contact with an immunoadsorbent comprised of a solid particulate substrate having covalently bound thereto antibodies specific to the antigen to form a bound and unbound fraction, said antibody including carboxyl and amine functional groups, said substrate including a barrier coating having attached thereto functional groups selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyazide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, said antibody being conjugated to said substrate by reaction between one of said carboxyl and amine groups with one of said functional groups attached to said substrate through said barrier coating, regenerating the immunoadsorbent by releasing substantially all of the antigens bound to the immunoadsorbent by rinsing said immunoadsorbent with an eluting solvent, detecting the labelled antigen in at least one or both of the unbound fraction and the bound fraction released by said eluting solvent, and reusing the immunoadsorbent for further assays of further samples containing the antigen to which said antibodies are specific.
Claim 2
Dependent Claim
236 words
In a method of radioimmunoassay wherein an antigen bearing sample and a radioactive labelled antigen are brought into contact with an immobilized immunoadsorbent to form a fraction bound to the immunoadsorbent and an unbound fraction and wherein the concentration of the antigen is determined as a function of the bound or unbound fraction, or both, the improvement comprising: flowing an antigen bearing sample and a radioactive labelled antigen into contact with an immunoadsorbent comprised of a solid particulate substrate having covalently bound thereto antibodies specific to the antigen to form a bound and unbound fraction, said antibody including carboxyl and amine functional groups, said substrate including a barrier coating having attached thereto functional groups selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyazide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, said antibody being conjugated to said substrate by reaction between one of said carboxyl and amine groups with one of said functional groups attached to said substrate through said barrier coating, regenerating the immunoadsorbent by releasing substantially all of the antigens bound to the immunoadsorbent by rinsing said immunoadsorbent with an eluting solvent, detecting the labelled antigen in at least one or both of the unbound fraction and the bound fraction released by said eluting solvent, and reusing the immunoadsorbent for further assays of further samples containing the antigen to which said antibodies are specific.
Claim 3
Dependent Claim
30 words
A method as set forth in claim 1 wherein said solid particulate substrate includes a particulate material composed of impervious glass spheres coated with an outer porous crust of microspheres.
Claim 4
Dependent Claim
33 words
A method as set forth in claim 1 wherein said particulate substrate includes a mass of porous refractory particles, and said barrier coating being a water insoluble material chemically bonded to said particles.
Claim 5
Dependent Claim
20 words
A method as set forth in claim 3 wherein said group is an azide and said barrier coating is dextran.
Claim 6
Dependent Claim
24 words
A method as set forth in claim 3 wherein said barrier coating is a polyolefin material and said functional group is a polyacyl azide.
Claim 7
Dependent Claim
23 words
A method as set forth in claim 3 wherein said barrier coating is a polyolefin material and said functional group is a polythiol.
Claim 8
Dependent Claim
33 words
A method as set forth in claim 3 wherein said barrier coating is a combination of a polyolefin material and a material selected from the group consisting of dextran, cellulose and carboxymethyl cellulose.
Claim 9
Dependent Claim
22 words
A method as set forth in claim 1 wherein said sample is preincubated with said antigen prior to flow through said immunoadsorbent.
Claim 10
Dependent Claim
55 words
A method as set forth in claim 1 wherein said substrate includes solid particulate material composed of impervious glass spheres each coated with an outer porous crust of microspheres, said spheres being of a diameter of between 5 and 500 microns, and said microspheres being of a diameter of between 5 millimicrons and 1 micron.
Claim 11
Dependent Claim
24 words
A method as set forth in claim 1 wherein said particulate substrate has a surface area of between 0.8 and 1.0 m 2 /gram.
Claim 12
Dependent Claim
28 words
A method as set forth in claim 1 wherein the activity of the immunoadsorbent is not less than 80% of the concentration of the antigen in the sample.
Claim 13
Independent Claim
99 words
A method of forming an immobilized immunoadsorbent for use in radioimmunoassay comprising: providing a particulate solid material having a porous surface and a barrier coating on said particulate solid, activating said barrier coating to attach thereto chemically a material having a functional group selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyacyl azide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, and conjugating to said barrier coated substrate an antibody specific to an antigen by covalently coupling through said groups to the carboxyl or amine group of said antibody.
Claim 14
Independent Claim
99 words
A method of forming an immobilized immunoadsorbent for use in radioimmunoassay comprising: providing a particulate solid material having a porous surface and a barrier coating on said particulate solid, activating said barrier coating to attach thereto chemically a material having a functional group selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyacyl azide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, and conjugating to said barrier coated substrate an antibody specific to an antigen by covalently coupling through said groups to the carboxyl or amine group of said antibody.
Claim 15
Dependent Claim
28 words
A method as set forth in claim 12 wherein said barrier coating is selected from the group consisting of polyolefin, dextran, carboxymethyl cellulose and cellulose, and combinations thereof.
Claim 16
Dependent Claim
16 words
A method as set forth in claim 12 wherein said functional group is an acyl azide.
Claim 17
Dependent Claim
16 words
A method as set forth in claim 12 wherein said functional group is a polyacyl azide.
Claim 18
Dependent Claim
15 words
A method as set forth in claim 12 wherein said functional group is a polythiol.
Claim 19
Dependent Claim
100 words
An immobilized immunoadsorbent for use in radioimmunoassay comprising: a solid particulate substrate having covalently bound thereto antibodies specific to an antigen, said antibody including carboxyl and amine functional groups, said substrate including a barrier coating having attached thereto functional groups selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyacyl azide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, and said antibody being conjugated to said substrate by reaction between one of said carboxyl and amine groups with one of said functional groups attached to said substrate through said barrier coating.
Claim 20
Dependent Claim
100 words
An immobilized immunoadsorbent for use in radioimmunoassay comprising: a solid particulate substrate having covalently bound thereto antibodies specific to an antigen, said antibody including carboxyl and amine functional groups, said substrate including a barrier coating having attached thereto functional groups selected from the group consisting of acyl azide, chlorothioformate, epoxide, carbonate, thiocarbonate, isocyanate, cyanuric chloride, polyacyl azide, polychlorothioformate, polyepoxide, polycarbonate, polythiocarbonate, polyisocyanate, polycyanuric chloride, polyimidocarbonate, polyaldehyde and polythiol, and said antibody being conjugated to said substrate by reaction between one of said carboxyl and amine groups with one of said functional groups attached to said substrate through said barrier coating.
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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) | 1981-08-21 | 1982-02-20 | 1982-08-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-08-21 | 1986-02-20 | 1986-08-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-08-21 | 1990-02-20 | 1990-08-22 | $7700.00 | Window Closed / Lapsed |