Patent US5071909A successfully refreshed from Google Patents.
US5071909A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Immobilization of proteins and peptides on insoluble supports
Assignee: Applied Biosystems LLC
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Filing: 1989-07-26
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Granted: 1991-12-10
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Expires: 2009-07-26
Abstract
Abstract The invention pertains to a method for immobilizing proteins or peptides onto a flat, microporous membrane surface in a form suitable for sequence analysis or other chemical or enzymatic processes. The process involves the formation of a thin polymer network that entraps the protein or peptide therein.
Inventors
- 👤 Darryl J. C. Pappin
- 👤 James M. Coull
- 👤 Hubert Koester
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 10:56
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/544; G01N33/545; B; B01; B01D; B01D69/00; B01D69/14; B01D69/141; C; C07; C07K; C07K1/00; C07K1/12; C07K1/128; C07K17/00; C07K17/02; C07K17/04; G01N33/54353; G01N33/5436; G01N33/68; G01N33/6803; G01N33/6818; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/815; Y10S530/817
Claims Analysis
Total Claims: 35
Claim 1
Independent Claim
92 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto the microporous membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; and c) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 2
Independent Claim
92 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto the microporous membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; and c) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 3
Dependent Claim
27 words
The method of claim 1, further comprising crosslinking the peptide or protein to the polymer network to thereby entrap the peptide or protein by covalent linkage thereto.
Claim 4
Dependent Claim
17 words
The method of claim 1, wherein the peptide or protein is physically entrapped within the polymer network.
Claim 5
Dependent Claim
35 words
The method of claim 1, wherein the microporous membrane surface is selected from the group consisting of teflon, polyvinylidene difluoride, nylon, cellulose acetate, nitrocellulose, polyester, polycarbonate, polysulphone, polystyrene, metals, zeolites, paper, silica, alumina and glass.
Claim 6
Dependent Claim
13 words
The method of claim 4, wherein the microporous membrane surface is polyvinylidene difluoride.
Claim 7
Dependent Claim
23 words
The method of claim 1, wherein the polymer is selected from the group consisting of polyethyleneimine, polyvinylamine, polyallylamine, polyacrylic acid and polymethacrylic acid.
Claim 8
Dependent Claim
17 words
The method of claim 1, wherein the polymer is crosslinked by a crosslinking agent or by photoactivation.
Claim 9
Dependent Claim
21 words
The method of claim 7, wherein the crosslinking agent is selected from the group consisting of methylenedianiline, ethylenedianiline, diaminostilbene and 1,4-phenylenediisothiocyanate.
Claim 10
Dependent Claim
12 words
The method of claim 7, wherein the photoactivation is by UV radiation.
Claim 11
Dependent Claim
25 words
The method of claim 1, wherein the microporous membrane surface is coated with a crosslinking agent before adsorbing the peptide or protein and polymer thereon.
Claim 12
Independent Claim
97 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polyamine polymer onto the adsorbed peptide or protein, wherein the polyamine polymer has functional groups which can be crosslinked; and c) crosslinking the polyamine polymer with 1,4-phenylenediisothiocyanate, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 13
Independent Claim
97 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polyamine polymer onto the adsorbed peptide or protein, wherein the polyamine polymer has functional groups which can be crosslinked; and c) crosslinking the polyamine polymer with 1,4-phenylenediisothiocyanate, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 14
Dependent Claim
27 words
The method of claim 11, further comprising crosslinking the peptide or protein to the polymer network to thereby entrap the peptide or protein by covalent linkage thereto.
Claim 15
Dependent Claim
17 words
The method of claim 11, wherein the peptide or protein is physically entrapped within the polymer network.
Claim 16
Dependent Claim
22 words
The method of claim 11, wherein the microporous membrane is coated with 1,4-phenylenediisothiocyanate before adsorbing the peptide or protein and polymer thereon.
Claim 17
Dependent Claim
11 words
The method of claim 11, wherein the polyamine polymer is polyallylamine.
Claim 18
Independent Claim
104 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polycarboxylic acid polymer onto the adsorbed peptide or protein, wherein the polycarboxylic acid polymer has functional groups which can be crosslinked; and c) crosslinking the polycarboxylic acid polymer with a dianiline crosslinking agent, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 19
Independent Claim
104 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing a polycarboxylic acid polymer onto the adsorbed peptide or protein, wherein the polycarboxylic acid polymer has functional groups which can be crosslinked; and c) crosslinking the polycarboxylic acid polymer with a dianiline crosslinking agent, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 20
Dependent Claim
27 words
The method of claim 16, further comprising crosslinking the peptide or protein to the polymer network to thereby entrap the peptide or protein by covalent linkage thereto.
Claim 21
Dependent Claim
17 words
The method of claim 16, wherein the peptide or protein is physically entrapped within the polymer network.
Claim 22
Dependent Claim
25 words
The method of claim 16, wherein the microporous membrane is coated with the dianiline crosslinking agent before adsorbing the peptide or protein and polymer thereto.
Claim 23
Dependent Claim
16 words
The method of claim 16, wherein the polycarboxylic acid polymer is polyacrylic acid or polymethacrylic acid.
Claim 24
Independent Claim
107 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing an azidopolymer or benzophenone polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; and c) crosslinking the azidopolymer or benzophenone polymer by exposing the microporous membrane to ultraviolet radiation, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 25
Independent Claim
107 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein onto a polyvinylidene difluoride membrane surface to noncovalently attach the peptide or protein thereto; b) depositing an azidopolymer or benzophenone polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; and c) crosslinking the azidopolymer or benzophenone polymer by exposing the microporous membrane to ultraviolet radiation, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 26
Dependent Claim
27 words
The method of claim 21, further comprising crosslinking the peptide or protein to the polymer network to thereby entrap the peptide or protein by covalent linkage thereto.
Claim 27
Dependent Claim
17 words
The method of claim 21, wherein the peptide or protein is physically entrapped within the polymer network.
Claim 28
Independent Claim
130 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein and a polymer onto the microporous membrane surface to noncovalently attach the peptide or protein thereto; b) treating the adsorbed peptide or protein with phenylisothiocyanate (PITC) and subsequently treating the adsorbed protein or peptide with a base; c) treating the product of step (b) with 1,4-phenylene diisothiocyanate (DITC); d) depositing a polymer onto the adsorbed peptide or protein of step (c), wherein the polymer has functional groups which can be crosslinked; and e) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 29
Independent Claim
130 words
A method of immobilizing a peptide or protein onto a flat, microporous membrane surface, comprising the steps of: a) adsorbing a peptide or protein and a polymer onto the microporous membrane surface to noncovalently attach the peptide or protein thereto; b) treating the adsorbed peptide or protein with phenylisothiocyanate (PITC) and subsequently treating the adsorbed protein or peptide with a base; c) treating the product of step (b) with 1,4-phenylene diisothiocyanate (DITC); d) depositing a polymer onto the adsorbed peptide or protein of step (c), wherein the polymer has functional groups which can be crosslinked; and e) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein, wherein the network does not interfere with subsequent chemical or enzymatic analysis of the protein or peptide.
Claim 30
Dependent Claim
35 words
The method of claim 24, wherein the microporous membrane surface is selected from the group consisting of teflon, polyvinylidene difluoride, nylon, cellulose acetate, nitrocellulose, polyester, polycarbonate, polysulphone, polystyrene, metals, zeolites, paper, silica, alumina and glass.
Claim 31
Dependent Claim
13 words
The method of claim 25, wherein the microporous membrane surface is polyvinylidene difluoride.
Claim 32
Dependent Claim
23 words
The method of claim 24, wherein the polymer is selected from the group consisting of polyethyleneimine, polyvinylamine, polyallylamine, polyacrylic acid and polymethacrylic acid.
Claim 33
Dependent Claim
17 words
The method of claim 24, wherein the polymer is crosslinked by a crosslinking agent or by photoactivation.
Claim 34
Dependent Claim
21 words
The method of claim 28, wherein the crosslinking agent is selected from the group consisting of methylenedianiline, ethylenedianiline, diaminostilbene and 1,4-phenylenediisothiocyanate.
Claim 35
Dependent Claim
25 words
The method of claim 24, wherein the microporous membrane surface is coated with a crosslinking agent before adsorbing the peptide or protein and polymer thereon.
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) | 1994-12-09 | 1995-06-10 | 1995-12-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-12-09 | 1999-06-10 | 1999-12-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-12-09 | 2003-06-10 | 2003-12-10 | $7700.00 | Window Closed / Lapsed |