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US5492840A
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Surface plasmon resonance sensor unit and its use in biosensor systems
Assignee: Cytiva Sweden AB
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Filing: 1989-11-09
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Granted: 1996-02-20
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Expires: 2009-11-09
Abstract
Abstract Methods for functionalizing sensing surfaces to be used in systems for measuring simultaneously several properties of one biomolecule as well as for simultaneously measuring the concentrations of a plurality of biomolecules in a sample. The invention furthermore also relates to sensor units containing such surfaces, to the use thereof for surface characterization of biomolecules, and to a reagent kit for functionalization of sensing surfaces.
Inventors
- 👤 Magnus Malmqvist
- 👤 Robert Karlsson
- 👤 Inger Ronnberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 18:45
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/17; G01N21/55; G01N21/552; G01N21/553; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54366; G01N33/54373; Y; Y10; Y10S; Y10S435/00; Y10S435/808; Y10S436/00; Y10S436/805; Y10S436/819
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
209 words
A sensor unit for use in a biosensor system based on surface plasmon resonance technology, comprising: a dielectric substrate which is replaceable in said biosensor system and which is coated with a metal film, said film having two or more sensing areas arranged so as to be passed in series or in parallel by a liquid stream, each of said sensing areas comprising a layer of an organic polymer or hydrogel over said metal film, containing at least one functional group supporting bi- or polyfunctional molecules capable of specific binding with biomolecules present in said liquid stream which are to be detected, said bi- or polyfunctional molecules comprising ligands that can specifically bind to biomolecules in a sample and a functional structure which can form reversible bonds with said at least one functional group which, through said functional structure, are coupled to said sensing area by a reversible bond, such that said two or more sensing areas can be regenerated for refunctionalization with the same or other bi- or polyfunctional molecules, wherein coupling between said bi- or polyfunctional molecules and said biomolecules in said liquid stream is selectively reversible such that the bi- or polyfunctional molecule-functionalized sensing areas can be regenerated, permitting repeated use of the functionalized sensor unit.
Claim 2
Dependent Claim
209 words
A sensor unit for use in a biosensor system based on surface plasmon resonance technology, comprising: a dielectric substrate which is replaceable in said biosensor system and which is coated with a metal film, said film having two or more sensing areas arranged so as to be passed in series or in parallel by a liquid stream, each of said sensing areas comprising a layer of an organic polymer or hydrogel over said metal film, containing at least one functional group supporting bi- or polyfunctional molecules capable of specific binding with biomolecules present in said liquid stream which are to be detected, said bi- or polyfunctional molecules comprising ligands that can specifically bind to biomolecules in a sample and a functional structure which can form reversible bonds with said at least one functional group which, through said functional structure, are coupled to said sensing area by a reversible bond, such that said two or more sensing areas can be regenerated for refunctionalization with the same or other bi- or polyfunctional molecules, wherein coupling between said bi- or polyfunctional molecules and said biomolecules in said liquid stream is selectively reversible such that the bi- or polyfunctional molecule-functionalized sensing areas can be regenerated, permitting repeated use of the functionalized sensor unit.
Claim 3
Dependent Claim
34 words
The sensor unit of claim 1, wherein said bi - or polyfunctional molecules are selected from a group of chimeric molecules having an identical sensing area binding structure, but having varying analyte binding structures.
Claim 4
Dependent Claim
17 words
The sensor unit of claim 1, wherein said metal film is a film of silver or gold.
Claim 5
Dependent Claim
30 words
The sensor unit of claim 1, wherein said sensing areas contain at least one functional group selected from the group consisting of hydroxyl, carboxyl, amino, aldehyde, hydrazide, carbonyl, and vinyl.
Claim 6
Dependent Claim
18 words
The sensor unit of claim 1, wherein a hydrogel is bound to said metal film via disulfide bonds.
Claim 7
Dependent Claim
17 words
The sensor unit of claim 1, wherein said hydrogel is a polysaccharide or a water-swellable organic polymer.
Claim 8
Dependent Claim
26 words
The sensor unit of claim 6 wherein said water-swellable organic polymer is selected from the group consisting of polyvinyl alcohol, polyacrylic acid, polyacrylamide, and polyethylene glycol.
Claim 9
Dependent Claim
32 words
The sensor unit of claim 6, wherein said polysaccharide is selected from the group consisting of agarose, dextran, carrageenan, alginic acid, starch, cellulose, and a derivative of any one of the foregoing.
Claim 10
Dependent Claim
13 words
The sensor unit of claim 8, wherein said derivative is a carboxymethyl derivative.
Claim 11
Dependent Claim
11 words
The sensor unit of claim 8, wherein said polysaccharide is dextran.
Claim 12
Dependent Claim
37 words
The sensor unit of claim 10, wherein said dextran is activated for binding said ligand or ligands by generating at least one member selected from the group consisting of carboxymethyl, carboxyl, hydrazide, ester, and disulfide groups therein.
Claim 13
Dependent Claim
15 words
The sensor unit of claim 11, wherein said ester groups are derivatized with a hapten.
Claim 14
Dependent Claim
11 words
The sensor unit of claim 12, wherein said hapten is theophylline.
Claim 15
Dependent Claim
28 words
The sensor unit of claim 1, wherein said functional group is selected from the group consisting of ion exchanger groups, metal chelating groups, and receptors for biological molecules.
Claim 16
Dependent Claim
17 words
The sensor unit of claim 1, wherein said metal film is coated with a layer of dextran.
Claim 17
Dependent Claim
19 words
The sensor unit of claim 15, wherein said chimeric molecules are antibodies against dextran conjugated with a biospecific ligand.
Claim 18
Dependent Claim
13 words
The sensor unit of claim 16, wherein said biospecific ligand is an immunoglobulin.
Claim 19
Dependent Claim
25 words
The sensor unit of claim 1, wherein said functional group is a molecule selected from the group consisting of theophylline, a steroid hormone, and thyroxine.
Claim 20
Dependent Claim
60 words
The sensor unit of claim 1, wherein said bi- or polyfunctional molecules contain a function for immobilizing said bi- or polyfunctional molecules on the corresponding sensing area, which function is common to all sensing areas, plus one or more analyte binding structures for interacting with biomolecules in said liquid stream, wherein said analyte binding structures vary between said sensing areas.
Claim 21
Dependent Claim
43 words
A method for detecting binding sites on a biomolecule, comprising: contacting a sample with said sensing areas of claim 1, and detecting any binding interacting between said biomolecule and said sensing areas, indicating the presence of a corresponding binding site on said biomolecule.
Claim 22
Dependent Claim
45 words
The method of claim 20, which further comprises providing a plurality of sensing areas, each having its own specific ligand, and serially or in parallel contacting a sample solution containing said biomolecule with said sensing areas in order to detect binding sites on said biomolecule.
Claim 23
Dependent Claim
152 words
A process for functionalizing said sensing areas of the sensor unit according to claim 1, comprising: introducing said sensor unit into a biosensor system based on surface plasmon technology which comprises a liquid handling block unit for conveying sample solutions to said sensing areas, and distributing by means of said liquid handling block unit a flow of a reagent solution containing a bi- or polyfunctional molecule to each of said sensing areas so that said bi- or polyfunctional molecule reversibly binds to said at least one functional group of said sensing areas, said bi- or polyfunctional molecule including (i) a functional group that binds to said sensing areas to immobilize said bi- or polyfunctional molecule on said sensing areas and (ii) at least one additional functional group that possesses specific binding affinity for a target molecule and which is exposed on each of said sensing areas for interaction with said target molecule.
Claim 24
Dependent Claim
33 words
The process of claim 22, wherein each of said sensing areas containing a unique functional group is arranged in series in said flow containing a bi- or polyfunctional ligand for each respective area.
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| 3.5 Years (Year 4) | 1999-02-19 | 1999-08-21 | 2000-02-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-02-19 | 2003-08-21 | 2004-02-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-02-19 | 2007-08-21 | 2008-02-20 | $7700.00 | Window Closed / Lapsed |