EP0410323A2
Expired / Lapsed
EP
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Immobilization of proteins and peptides on insoluble supports
Assignee: Applied Biosystems LLC
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Filing: 1990-07-20
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Granted: —
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Expires: 2010-07-20
Abstract
Abstract The invention pertains to a method for immobilizing proteins or pep tides 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
EP
Data Source
google_patents
Last Synchronized
2026-11-05 21:10
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: 27
Claim 1
Independent Claim
60 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, wherein the polymer has functional groups which can be crosslinked; and b) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein.
Claim 2
Independent Claim
60 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, wherein the polymer has functional groups which can be crosslinked; and b) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein.
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
20 words
The method of Claim 1, wherein the peptide or protein and the polymer are simultaneously adsorbed onto the microporous membrane.
Claim 6
Dependent Claim
20 words
The method of Claim 1, wherein the peptide or protein and the polymer are sequentially adsorbed onto the microporous membrane.
Claim 7
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 8
Dependent Claim
13 words
The method of Claim 6, wherein the microporous membrane surface is polyvinylidene difluoride.
Claim 9
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 10
Dependent Claim
17 words
The method of Claim 1, wherein the polymer is crosslinked by a crosslinking agent or by photoactivation.
Claim 11
Dependent Claim
21 words
The method of Claim 9, wherein the crosslinking agent is selected from the group consisting of methylenedianiline, ethylenedianiline, diaminostilbene and 1,4-phenylenediisothiocyanate.
Claim 12
Dependent Claim
12 words
The method of Claim 9, wherein the photoactivation is by UV radiation.
Claim 13
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 14
Independent Claim
96 words
A method for determining the amino acid sequence of a peptide or protein, comprising the steps of: a) transferring a peptide or protein from a separatory gel onto a flat, microporous membrane under conditions such that the peptide or protein is adsorbed onto the microporous membrane; b) depositing a polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; c) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein; and d) determining the amino acid sequence of the entrapped peptide or protein.
Claim 15
Independent Claim
96 words
A method for determining the amino acid sequence of a peptide or protein, comprising the steps of: a) transferring a peptide or protein from a separatory gel onto a flat, microporous membrane under conditions such that the peptide or protein is adsorbed onto the microporous membrane; b) depositing a polymer onto the adsorbed peptide or protein, wherein the polymer has functional groups which can be crosslinked; c) crosslinking the polymer, to thereby produce a polymer network which entraps the peptide or protein therein; and d) determining the amino acid sequence of the entrapped peptide or protein.
Claim 16
Dependent Claim
22 words
The method of Claim 13, wherein the peptide or protein is transferred from the separatory gel onto the microporous membrane by electroblotting.
Claim 17
Dependent Claim
18 words
The method of Claim 13 wherein the peptide or protein is sequenced using solid-phase Edman protein sequencing methods.
Claim 18
Dependent Claim
34 words
The method of Claim 13, wherein 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 19
Dependent Claim
23 words
The method of Claim 13, wherein the polymer is selected from the group consisting of polyethyleneimine, polyvinylamine, polyallylamine, polyacrylic acid and polymethacrylic acid.
Claim 20
Dependent Claim
17 words
The method of Claim 13, wherein the polymer is crosslinked by a crosslinking agent or by photoactivation.
Claim 21
Dependent Claim
21 words
The method of Claim 18, wherein the crosslinking agent is selected from the group consisting of methylenedianiline, ethylenedianiline, diaminostilbene and 1,4-phenylenediisothiocyanate.
Claim 22
Dependent Claim
38 words
A microporous membrane having a peptide or protein immobilized on its surface, comprising: a) a flat, microporous membrane; b) a peptide or protein adsorbed thereon; and c) a crosslinked polymer network which entraps the peptide or protein therein.
Claim 23
Dependent Claim
38 words
A microporous membrane having a peptide or protein immobilized on its surface, comprising: a) a flat, microporous membrane; b) a peptide or protein adsorbed thereon; and c) a crosslinked polymer network which entraps the peptide or protein therein.
Claim 24
Dependent Claim
19 words
The microporous membrane of Claim 20 wherein the polymer network entraps the peptide or protein by covalent linkage thereto.
Claim 25
Dependent Claim
18 words
The microporous membrane of Claim 20, wherein the peptide or protein is physically entrapped within the polymer network.
Claim 26
Dependent Claim
35 words
The microporous membrane of Claim 20, wherein the microporous 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 27
Dependent Claim
24 words
The microporous membrane of Claim 20 wherein the polymer is selected from the group consisting of polyethyleneimine, polyvinylamine, polyallylamine, polyacrylic acid and polymethacrylic acid.
References Cited (Backward Citations)
No citation records indexed.
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