US5661019A
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US
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Trifunctional conjugates
Assignee: Beckman Coulter Inc
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Filing: 1995-03-22
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Granted: 1997-08-26
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Expires: 2015-03-22
Abstract
Abstract A trifunctional conjugate is providing having three chemical moieties attached through a spacer moiety. At least two of the chemical moieties are relatively small molecules, usually less than about 7,000 Daltons in size. The spacer moiety is selected to impart certain steric properties to the conjugate. In one embodiment, the binding of a macromolecular specific binding partner to one of the chemical moieties sterically inhibits the binding of a different macromolecule to another chemical moieties. In another embodiment, the binding of a first chemical moiety to a macromolecule restricts the subsequent binding of a second tridentate member to a proximate location on the same macromolecule. The three chemical moieties are preferably a nitrophenylazido residue, a phenyl boronic acid residue, and a solid support or a label such as biotin. The spacer is preferably cysteine, lysine, glutamic acid, pyroglutamic acid, S-acetylmercaptosuccinic anhydride or ω-carbobenzoxylysine. The conjugate is useful in immunoassays and for targeted labeling of proteins.
Inventors
- 👤 Chan S. Oh
- 👤 James C. Sternberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-10 02:36
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54353; G01N33/531; G01N33/532; G01N33/536; G01N33/537; G01N33/539; G01N33/54313; G01N33/94; Y; Y10; Y10S; Y10S435/00; Y10S435/964; Y10S436/00; Y10S436/819; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815; Y10S530/816
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
179 words
A trifunctional conjugate having a first member, a second member, and a third member, each member being attached to an organic spacer moiety, the organic spacer moiety being attached to all three members so that the first member is attached to the second member through covalent bonds, the first member is attached to the third member through covalent bonds, and the second member is attached to the third member through covalent bonds, wherein: a) the first member is a small molecule capable of noncovalently binding to a first member binding partner; b) the second member is a modulating member capable of noncovalently binding to a modulating member specific binding partner; c) the third member is a small molecule capable of binding to a third member binding partner; and d) the organic spacer moiety is selected such that the binding of the modulating member to the modulating member binding partner prevents, by steric inhibition, at least one of the first member binding partner or the third member binding partner from binding to the first member or the third member, respectively.
Claim 2
Dependent Claim
179 words
A trifunctional conjugate having a first member, a second member, and a third member, each member being attached to an organic spacer moiety, the organic spacer moiety being attached to all three members so that the first member is attached to the second member through covalent bonds, the first member is attached to the third member through covalent bonds, and the second member is attached to the third member through covalent bonds, wherein: a) the first member is a small molecule capable of noncovalently binding to a first member binding partner; b) the second member is a modulating member capable of noncovalently binding to a modulating member specific binding partner; c) the third member is a small molecule capable of binding to a third member binding partner; and d) the organic spacer moiety is selected such that the binding of the modulating member to the modulating member binding partner prevents, by steric inhibition, at least one of the first member binding partner or the third member binding partner from binding to the first member or the third member, respectively.
Claim 3
Dependent Claim
28 words
The trifunctional conjugate of claim 1 wherein said organic spacer moiety is selected from the group consisting of cysteine, lysine, glutamic acid, pyroglutamic acid, S-acetylmercaptosuccinic anhydride, and ω-carbobenzoxylysine.
Claim 4
Dependent Claim
42 words
The trifunctional conjugate of claim 1 wherein the distance between the first and second members, the distance between the first and third members, and the distance between the second and third members are each from about 18 Å to about 70 Å.
Claim 5
Dependent Claim
52 words
A preparation comprising at least two trifunctional conjugate molecules of claim 1 wherein the first member, the modulating member and the third member of the conjugates are separated from each other by distances controlled to within about one bond length, the controlled distances being substantially uniform for the molecules of the preparation.
Claim 6
Dependent Claim
22 words
The trifunctional conjugate of claim 1 wherein the first member, the modulating member and the third member are each small molecule ligands.
Claim 7
Dependent Claim
20 words
The trifunctional conjugate of claim 1 wherein the modulating member is selected from the group consisting of theophylline and theophylline-amine.
Claim 8
Dependent Claim
20 words
The trifunctional conjugate of claim 1 wherein the first member is selected from the group consisting of biotin and DNP.
Claim 9
Dependent Claim
20 words
The trifunctional conjugate of claim 1 wherein the third member is selected from the group consisting of biotin and DNP.
Claim 10
Dependent Claim
26 words
The trifunctional conjugate of claim 1 wherein said organic spacer moiety is selected from the group consisting of cysteine, lysine, glutamic acid, pyroglutamic acid and carbobenzoxylysine.
Claim 11
Dependent Claim
68 words
The trifunctional conjugate of claim 1 wherein the organic spacer moiety is selected such that the binding of a first macromolecule, having at least one first proximity label conjugated thereto, to the first member, and the binding of a second macromolecule, having at least one second proximity label conjugated thereto, to the third member, produces a measurable reaction between the first proximity label and the second proximity label.
Claim 12
Dependent Claim
15 words
The trifunctional conjugate of claim 10 wherein the first and second proximity labels are enzymes.
Claim 13
Dependent Claim
25 words
The trifunctional conjugate of claim 10 wherein the first and second proximity labels are selected from the group consisting of energy donors and energy acceptors.
Claim 14
Dependent Claim
197 words
A trifunctional conjugate having a first member, a second member, and a third member, each member being attached to an organic spacer moiety, the organic spacer moiety being attached to all three members so that the first member is attached to the second member through covalent bonds, the first member is attached to the third member through covalent bonds, and the second member is attached to the third member through covalent bonds, wherein: a) the first member is a small molecule ligand capable of noncovalently binding to a small molecule ligand binding partner having at least one proximity label conjugated thereto; b) the second member is a modulating member capable of noncovalently binding to a modulating member specific binding partner; c) the third member is selected from the group consisting of: i) at least one second proximity label, and ii) at least one solid support having at least one second proximity label conjugated thereto; and d) the organic spacer moiety is selected such that the binding of the modulating member to the modulating member specific binding partner prevents, by steric inhibition, the generation of a measurable reaction between said first proximity label and said second proximity label.
Claim 15
Dependent Claim
197 words
A trifunctional conjugate having a first member, a second member, and a third member, each member being attached to an organic spacer moiety, the organic spacer moiety being attached to all three members so that the first member is attached to the second member through covalent bonds, the first member is attached to the third member through covalent bonds, and the second member is attached to the third member through covalent bonds, wherein: a) the first member is a small molecule ligand capable of noncovalently binding to a small molecule ligand binding partner having at least one proximity label conjugated thereto; b) the second member is a modulating member capable of noncovalently binding to a modulating member specific binding partner; c) the third member is selected from the group consisting of: i) at least one second proximity label, and ii) at least one solid support having at least one second proximity label conjugated thereto; and d) the organic spacer moiety is selected such that the binding of the modulating member to the modulating member specific binding partner prevents, by steric inhibition, the generation of a measurable reaction between said first proximity label and said second proximity label.
References Cited (Backward Citations)
US4067774A
Cited
US3935074A
Cited
US4134792A
Cited
US4130462A
Cited
US4243749A
Cited
US4233402A
Cited
US4298685A
Cited
US4238565A
Cited
GB2029011A
Cited
US4228237A
Cited
GB2084317A
Cited
EP0055869A2
Cited
US4711955A
Cited
US4604365A
Cited
EP0070685A2
Cited
US4608336A
Cited
US4629690A
Cited
US4506009A
Cited
US4663278A
Cited
US4722906A
Cited
US4687735A
Cited
US4687732A
Cited
US4550075A
Cited
US4582789A
Cited
EP0177191A1
Cited
EP0183901A2
Cited
US4760142A
Cited
US4868104A
Cited
EP0220899A1
Cited
US4680338A
Cited
US4791067A
Cited
US5168057A
Cited
US5196351A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2000-08-25 | 2001-02-24 | 2001-08-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2004-08-25 | 2005-02-24 | 2005-08-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2008-08-25 | 2009-02-24 | 2009-08-26 | $7700.00 | Window Closed / Lapsed |