US5244785A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Determination of high molecular weight analytes using a β-galactosidase complementation assay
Assignee: Microgenics Corp
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Filing: 1991-02-01
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Granted: 1993-09-14
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Expires: 2011-02-01
Abstract
Abstract A method for detecting the presence of a high-molecular-weight analyte, which uses a variation on complementation assays, comprising (1) providing an enzyme donor (ED)complex comprising ED coupled to an analyte-specific binding molecule, wherein the ED complex retains measurable complementation activity such that active β-galactosidase is formed in the presence of an enzyme acceptor (EA); (2) contacting the ED complex and EA in the presence of a sample suspected of containing a high-molecular-weight analyte that reacts specifically with the analyte-specific binding molecule; and (3) relating the presence of the analyte in the sample to the formation of active β-galactosidase enzyme.
Inventors
- 👤 Rueyming Loor
- 👤 Jeff Shindelman
- 👤 Pyare L. Khanna
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 21:35
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/581; G01N33/53; G01N33/536; G01N33/542; Y; Y10; Y10S; Y10S435/00; Y10S435/975
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
101 words
A method for detecting the presence of an analyte having a molecular weight of at least 10,000 daltons using a β-galactosidase complementation assay, comprising: (a) providing an enzyme donor (ED) complex comprising ED coupled to a binding moiety that reacts specifically with said analyte, wherein said D complex retains measurable complementation activity such that active β-galactosidase is formed in the present of enzyme acceptor (EA); (b) contacting the ED complex and EA in the presence of a sample suspected of containing said analyte, and (c) relating the presence of the analyte in the sample to the formation of active β-galactoside enzyme.
Claim 2
Independent Claim
101 words
A method for detecting the presence of an analyte having a molecular weight of at least 10,000 daltons using a β-galactosidase complementation assay, comprising: (a) providing an enzyme donor (ED) complex comprising ED coupled to a binding moiety that reacts specifically with said analyte, wherein said D complex retains measurable complementation activity such that active β-galactosidase is formed in the present of enzyme acceptor (EA); (b) contacting the ED complex and EA in the presence of a sample suspected of containing said analyte, and (c) relating the presence of the analyte in the sample to the formation of active β-galactoside enzyme.
Claim 3
Dependent Claim
71 words
A method according to claim 1, wherein said ED complex, EA, and sample are contacted with an enzyme substrate which provides a measurable product upon reaction with β-galactosidase, the measurable product provided by the reaction of the enzyme substrate and active enzyme is measured, and the amount of said analyte in the sample is determined by comparing the amount of measurable product to that formed from a known amount of analyte.
Claim 4
Dependent Claim
14 words
A method according to claim 2, wherein said enzyme substrate is chlorophenol red β-galactoside.
Claim 5
Dependent Claim
34 words
A method according to claim 1, wherein excess unlabelled polyclonal or monoclonal antibody specific for said analyte s also present when said ED complex is contacted with EA in the present of said analyte.
Claim 6
Dependent Claim
12 words
A method according to claim 1, wherein said analyte is a polypeptide.
Claim 7
Dependent Claim
18 words
A method according to claim 1, wherein said analyte has a molecular weight of at least 20,000 daltons.
Claim 8
Dependent Claim
15 words
A method according to claim 6, wherein id binding moiety is a DNA or RNA.
Claim 9
Dependent Claim
13 words
A method according to claim 1, wherein said analyte is a nucleic acid.
Claim 10
Dependent Claim
12 words
A method according to claim 8, wherein id nucleic acid is DNA.
Claim 11
Dependent Claim
12 words
A method according to claim 8, wherein said nucleic acid is RNA.
Claim 12
Dependent Claim
13 words
A method according to claim 1, wherein said analyte is a virus particle.
Claim 13
Dependent Claim
17 words
A method according to claim 1, wherein said binding moiety is an antibody or a natural receptor.
Claim 14
Dependent Claim
21 words
A method according to claim 12, wherein said antibody is a monoclonal antibody or recombinant peptide fragment equivalent of an antibody.
Claim 15
Dependent Claim
24 words
A method according to claim 1, wherein said ED is linked to an antibody specific for said analyte by a bifunctional organic linking group.
Claim 16
Dependent Claim
15 words
A method according to claim 1, wherein said ED complex is a recombinant fusion protein.
References Cited (Backward Citations)
US4592998A
Cited
WO1986002666A1
Cited
US4708929A
Cited
WO1989002597A1
Cited
US4994385A
Cited
US5037735A
Cited
WO1990013569A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-09-13 | 1997-03-15 | 1997-09-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-09-13 | 2001-03-15 | 2001-09-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-09-13 | 2005-03-15 | 2005-09-14 | $7700.00 | Window Closed / Lapsed |