US4081246A
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Solid phase column immunoassay procedure employing novel immunochemical composites
Assignee: Beckman Coulter Inc
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Filing: 1976-05-03
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Granted: 1978-03-28
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Expires: 1996-05-03
Abstract
Abstract An improved method of separating free from bound fractions in a solid phase column immunoassay procedure of the type wherein a solution is contacted with a composite comprising an activated polysaccharide matrix covalently coupled to a primary antibody, wherein the improvements comprise: (a) coupling an α,ω-diaminospacer to said activated polysaccharide matrix via one of said α,ω-diamino-spacer's amino groups thereby forming a derivatized polysaccharide matrix; and (b) covalently coupling said derivatized polysaccharide matrix to an antibody selected from a group consisting of primary and secondary antibodies via a bifunctional coupling agent having a formula ##STR1## WHEREIN N IS AN INTEGER FROM 1 AND 6 AND WHEREIN E IS AN INTEGER FROM 1 TO 2.
Inventors
- 👤 Alan J. Polito
- 👤 William S. Knight
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 11:44
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/539; G01N33/541; A; A61; A61K; A61K39/00; A61K39/395; A61K39/44; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/813; Y10S530/815; Y10S530/816
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
117 words
A method of separating free from bound fractions in a solid phase column immunoassay procedure of the type wherein a solution is contacted with a composite comprising an activated polysaccharide matrix covalently coupled to a primary antibody, wherein the improvement comprises contacting said solution with a composite comprising a derivatized, polysaccharide matrix covalently coupled to an antibody selected from a group consisting of primary and secondary antibodies via a bifunctional coupling agent having a formula ##STR10## wherein n is an integer from 1 to 6 and wherein e is an integer from 1 to 2; and wherein said derivatized polysaccharide matrix comprises an activated, polysaccharide matrix coupled to an α,ω-diaminospacer via one of said α,ω-diaminospacer's amino groups.
Claim 2
Independent Claim
117 words
A method of separating free from bound fractions in a solid phase column immunoassay procedure of the type wherein a solution is contacted with a composite comprising an activated polysaccharide matrix covalently coupled to a primary antibody, wherein the improvement comprises contacting said solution with a composite comprising a derivatized, polysaccharide matrix covalently coupled to an antibody selected from a group consisting of primary and secondary antibodies via a bifunctional coupling agent having a formula ##STR10## wherein n is an integer from 1 to 6 and wherein e is an integer from 1 to 2; and wherein said derivatized polysaccharide matrix comprises an activated, polysaccharide matrix coupled to an α,ω-diaminospacer via one of said α,ω-diaminospacer's amino groups.
Claim 3
Dependent Claim
22 words
The method of claim 1 wherein said immunochemical composite has a formula ##STR11## wherein m is an integer from 1 to 12.
Claim 4
Dependent Claim
43 words
The method of claim 2 wherein said polysaccharide matrix is selected from a group consisting of cellulosic polymers, dextran polymers, agarose, and derivatives thereof, wherein m is an integer from 4 to 6, and wherein n is an integer from 4 to 6.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein said antibody is a secondary antibody.
Claim 6
Dependent Claim
33 words
The method of claim 4 wherein a primary immunoassay reaction is performed outside of said column and then an aliquot of said reaction mixture is incubated on said solid phase second antibody column.
Claim 7
Dependent Claim
13 words
The method of claim 5 wherein said immunoassay procedure is a radioimmunoassay procedure.
Claim 8
Dependent Claim
19 words
The method of claim 5 wherein an excess of said second antibody is present on said solid phase column.
Claim 9
Dependent Claim
13 words
The method of claim 7 wherein said immunoassay procedure is a radioimmunoassay procedure.
Claim 10
Dependent Claim
46 words
The method of claim 4 wherein said solid phase second antibody columns are loaded with primary antibody, labeled antigen, and sample, and wherein both said primary reaction and said incubation step with solid preparation of second antibody proceed simultaneously in the void volume of the column.
Claim 11
Dependent Claim
13 words
The method of claim 9 wherein said immunoassay procedure is a radioimmunoassay procedure.
Claim 12
Dependent Claim
36 words
The method of claim 4 wherein primary antibody is incubated on said solid phase second antibody column thereby producing a primary antibody column and wherein labeled antigen and sample are incubated on said primary antibody column.
Claim 13
Dependent Claim
13 words
The method of claim 11 wherein said immunoassay proceduere is a radioimmunoassay procedure.
Claim 14
Dependent Claim
13 words
The method of claim 1 wherein said immunoassay procedure is a radioimmunoassay procedure.
Claim 15
Dependent Claim
120 words
An immunochemical apparatus for separating free from bound fractions in a solid phase, immunoassay procedure of the type comprising a reaction vessel and a plurality of immunochemical composites, wherein each immunochemical composite comprises an activated polysaccharide matrix covalently coupled to a primary antibody, wherein the improvement comprises an immunochemical composite comprising a derivatized, polysaccharide matrix covalently coupled to an antibody selected from a group consisting of primary and secondary antibodies via a bifunctional coupling agent having a formula ##STR12## wherein n is an integer from 1 to 6 and wherein e is an integer from 1 to 2; and wherein said derivatized, polysaccharide matrix comprises an activated, polysaccharide matrix coupled to an α,ω-diaminospacer via one of said α,ω-diaminospacer's amino groups.
Claim 16
Dependent Claim
120 words
An immunochemical apparatus for separating free from bound fractions in a solid phase, immunoassay procedure of the type comprising a reaction vessel and a plurality of immunochemical composites, wherein each immunochemical composite comprises an activated polysaccharide matrix covalently coupled to a primary antibody, wherein the improvement comprises an immunochemical composite comprising a derivatized, polysaccharide matrix covalently coupled to an antibody selected from a group consisting of primary and secondary antibodies via a bifunctional coupling agent having a formula ##STR12## wherein n is an integer from 1 to 6 and wherein e is an integer from 1 to 2; and wherein said derivatized, polysaccharide matrix comprises an activated, polysaccharide matrix coupled to an α,ω-diaminospacer via one of said α,ω-diaminospacer's amino groups.
Claim 17
Dependent Claim
23 words
The immunochemical apparatus of claim 14 wherein said immunochemical composite has a formula ##STR13## wherein m is an integer from 1 to 12.
Claim 18
Dependent Claim
43 words
The immunochemical apparatus claim 15 wherein said polysaccharide matrix is selected from a group consisting of cellulosic polymers, dextran polymers, agarose, and derivatives thereof, wherein m is an integer from 4 to 6 and wherein n is an integer from 4 to 6.
Claim 19
Dependent Claim
13 words
The immunochemical apparatus of claim 16 wherein said antibody is a secondary antibody.
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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) | 1981-03-27 | 1981-09-26 | 1982-03-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-03-27 | 1985-09-26 | 1986-03-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-03-27 | 1989-09-26 | 1990-03-28 | $7700.00 | Window Closed / Lapsed |