US5077210A
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Immobilization of active agents on substrates with a silane and heterobifunctional crosslinking agent
Assignee: Science Applications International Corp SAIC
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Filing: 1989-01-13
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Granted: 1991-12-31
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Expires: 2009-01-13
Abstract
Abstract Active agents such as proteins are covalently immobilized on substrates carrying hydroxyl groups. A silane is bound to the substrate and coupled to a heterobifunctional crosslinker at one functional group leaving a free functional group, different than the first group, to which a protein is bound while retaining high protein functionality. Preferably, the silane has a functional group which reacts with the hydroxyl group of the substrate and a thiol terminal group which reacts with a functional group of a heterobifunctional crosslinking agent which contains a succinimide group that reacts with an amino group of the active agent. Bound active agents such as proteins are useful as biosensors or reactants in a variety of applications including bioassays.
Inventors
- 👤 Frances S. Eigler
- 👤 Jacque Georger
- 👤 Suresh K. Bhatia
- 👤 Jeff Calvert
- 👤 Lisa C. Shriver-Lake
- 👤 Reinhard Bredehorst
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 03:41
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/551; G01N33/552; C; C12; C12N; C12N11/00; C12N11/02; C12N11/06; C12N11/14; G01N33/54353; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/811; Y10S530/812; Y10S530/815; Y10S530/816
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
125 words
A method for modifying a substrate to activate said substrate for the immobilization of anzime group-containing active agents, said method comprising, selecting a substrate having hydroxyl groups on at least one surface, coating said hydroxyl-containing surface with a thiol-terminal group silane containing a reactive group other than a thiol group that reacts only with the hydroxyl groups on said surface to form a silane layer having an available reactive thiol group, coating said silane layer with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts only with the thiol-terminal group on said silane to form a layer of crosslinking agent having an available free succinimide reactive group capable of reacting with amino group of said active agent.
Claim 2
Independent Claim
125 words
A method for modifying a substrate to activate said substrate for the immobilization of anzime group-containing active agents, said method comprising, selecting a substrate having hydroxyl groups on at least one surface, coating said hydroxyl-containing surface with a thiol-terminal group silane containing a reactive group other than a thiol group that reacts only with the hydroxyl groups on said surface to form a silane layer having an available reactive thiol group, coating said silane layer with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts only with the thiol-terminal group on said silane to form a layer of crosslinking agent having an available free succinimide reactive group capable of reacting with amino group of said active agent.
Claim 3
Dependent Claim
39 words
A method in accordance with the method of claim 1 and further comprising, coating said layer of crosslinking agent with an active agent having an amine group that reacts with the available reactive succinimide group on the crosslinking agent.
Claim 4
Dependent Claim
12 words
A method in accordance with claim 1 wherein said silane is mercaptomethyldimethylethoxysilane.
Claim 5
Dependent Claim
12 words
A method in accordance with claim 1 wherein said silane is mercaptopropyltrimethoxysilane.
Claim 6
Dependent Claim
20 words
A method in accordance with the method of claim 1 wherein said heterobifunctional crosslinking agent is N gamma-maleimidobutyryloxy succinimide ester.
Claim 7
Dependent Claim
19 words
A method in accordance with the method of claim 1 wherein said heterobifunctional crosslinking agent is N-succinimidyl-3 (2-pyridyldithio) propionate.
Claim 8
Dependent Claim
19 words
A method in accordance with the method of claim 1 wherein said heterobifunctional crosslinking agent is N-succinimdyl (4-iodoacetyl) aminobenzoate.
Claim 9
Dependent Claim
19 words
A method in accordance with the method of claim 1 wherein said heterobifunctional crosslinking agent is succinimidyl 4-(p-maleimidophenyl) butyrate.
Claim 10
Dependent Claim
16 words
A method in accordance with claim 1 wherein said substrate is formed of an inorganic material.
Claim 11
Dependent Claim
19 words
A method in accordance with claim 1 wherein said succinimide group is on one end of said crosslinking agent.
Claim 12
Dependent Claim
16 words
A product for supporting an active agent amino group-containing produced by the method of claim 1.
Claim 13
Dependent Claim
32 words
A product in accordance with claim 11 and further comprising, a layer of amino group-containing active agent bound by said amino group to the free succinimide reactive group of said crosslinking agent.
Claim 14
Dependent Claim
12 words
The product of claim 11 wherein said substrate is an inorganic substrate.
Claim 15
Dependent Claim
10 words
The product of claim 11 wherein said silane is mercaptomethyldimethylethoxysilane.
Claim 16
Dependent Claim
10 words
The product of claim 11 wherein said silane is mercaptopropyltrimethoxysilane.
Claim 17
Dependent Claim
13 words
The product of claim 11 wherein said crosslinking agent is N-gamma-maleimidobutyryloxy succinimide ester.
Claim 18
Dependent Claim
14 words
The product of claim 11 wherein said crosslinking agent is N succinimidyl-3 (2-pyridyldithio) propionate.
Claim 19
Dependent Claim
11 words
The product of claim 11 wherein said crosslinking agent is N-Succinimdyl-(4-iodoacetyl)aminobenzoate.
Claim 20
Dependent Claim
13 words
The product of claim 11 wherein said crosslinking agent is succinimidyl 4-(p-maleimidophenyl) butyrate.
Claim 21
Dependent Claim
14 words
The product of claim 12 wherein said crosslinking agent is succinimidyl 4 (maleimidophenyl) butyrate.
Claim 22
Dependent Claim
17 words
A product of claim 11 wherein said succinimide group is on one end of said crosslinking agent.
Claim 23
Dependent Claim
119 words
In a method of immobilizing an amino group-containing active agent on a substrate, the improvement comprising preparing said substrate by, selecting a substrate having hydroxyl groups on a surface thereof, depositing on the substrate of film of a thiol-terminal group silane having a reactive group other than the thiol group that reacts with said hydroxyl groups to form silane surface having a free reactive thiol group, coating said silane surface with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts with said free reactive thiol group to form a layer of the crosslinking agent having an available free succinimide reactive group that reacts with the amino group of said active agent.
Claim 24
Dependent Claim
119 words
In a method of immobilizing an amino group-containing active agent on a substrate, the improvement comprising preparing said substrate by, selecting a substrate having hydroxyl groups on a surface thereof, depositing on the substrate of film of a thiol-terminal group silane having a reactive group other than the thiol group that reacts with said hydroxyl groups to form silane surface having a free reactive thiol group, coating said silane surface with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts with said free reactive thiol group to form a layer of the crosslinking agent having an available free succinimide reactive group that reacts with the amino group of said active agent.
Claim 25
Dependent Claim
32 words
A method in accordance with the method of claim 22 wherein said active agent is selected from the group consisting of enzymes, nucleic acid probes, lectins, chelators, ionophores, lipids, and biological cells.
Claim 26
Dependent Claim
12 words
A method in accordance with claim 23 herein said substrate is inorganic.
Claim 27
Dependent Claim
24 words
A method in accordance with the method of claim 22 wherein said active agent is selected from the group consisting of antibodies and antigens.
Claim 28
Dependent Claim
17 words
A method in accordance with the method of claim 22 wherein said active agent is a protein.
Claim 29
Dependent Claim
17 words
A method in accordance with the method of claim 22 wherein said active agent is a receptor.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1994-12-30 | 1995-07-01 | 1995-12-31 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-12-30 | 1999-07-01 | 1999-12-31 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-12-30 | 2003-07-01 | 2003-12-31 | $7700.00 | Window Closed / Lapsed |