US4923901A
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Membranes with bound oligonucleotides and peptides
Assignee: EMD Millipore Corp
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Filing: 1987-09-04
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Granted: 1990-05-08
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Expires: 2007-09-04
Abstract
Abstract A method is provided for synthesizing oligonucleotides and peptides directly onto a membrane. The method provides a means for generating membrane affinity supports. A modified membrane for the method of direct synthesis is also provided.
Inventors
- 👤 Hubert Koester
- 👤 James M. Coull
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 07:12
CPC Classifications
B; B01; B01D; B01D67/00; B01D67/0081; B01D67/0093; B01D67/00931; B01J; B01J19/00; B01J19/0046; C; C07; C07H; C07H21/00; C07K; C07K1/00; C07K1/04; C07K1/042; C07K17/00; C07K17/02; C07K17/06; B01D2323/00; B01D2323/30; B01D2323/38; B01J2219/00; B01J2219/00274; B01J2219/00277; B01J2219/00497; B01J2219/00527; B01J2219/00583; B01J2219/0059; B01J2219/00596; B01J2219/00603; B01J2219/00605; B01J2219/0061; B01J2219/00623; B01J2219/00626; B01J2219/00632; B01J2219/00637; B01J2219/00639; B01J2219/00641; B01J2219/00718; B01J2219/0072; B01J2219/00722; B01J2219/00725; C40; C40B; C40B40/00; C40B40/04; C40B40/06; C40B40/10; Y; Y10; Y10T; Y10T428/00; Y10T428/249921; Y10T428/249953; Y10T428/249954
Claims Analysis
Total Claims: 52
Claim 1
Independent Claim
16 words
A modified membrane comprising a polymeric membrane having a protected nucleoside or amino acid linked thereto.
Claim 2
Dependent Claim
16 words
A modified membrane comprising a polymeric membrane having a protected nucleoside or amino acid linked thereto.
Claim 3
Dependent Claim
85 words
A modified membrane represented by the formula: P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through its functional group Y to the functional group X; and S W is a protected nucleoside or amino acid, S W being bound to the linker through the functional group Z of the linker.
Claim 4
Dependent Claim
85 words
A modified membrane represented by the formula: P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through its functional group Y to the functional group X; and S W is a protected nucleoside or amino acid, S W being bound to the linker through the functional group Z of the linker.
Claim 5
Dependent Claim
34 words
A modified membrane of claim 2, wherein the membrane P comprises a flat, permeable polymeric material of porous structure which contains functional groups within its constituent monomeric units for attachment of the spacer group.
Claim 6
Dependent Claim
28 words
A modified membrane of claim 3, wherein the monomers are acrylic or methacrylic acid esters having a free alcohol or ester function for attachment of the spacer group.
Claim 7
Dependent Claim
27 words
A modified membrane of claim 3, wherein the polymeric material is a cross-linked polymer selected from the group consisting of polydialkylsilandiols, polydialkylsiloxanes, polyvinyl alcohols, polyoxymethylenes and polyoxyethylenes.
Claim 8
Dependent Claim
31 words
A modified membrane of claim 2, wherein the membrane is a flat, permeable, polymeric material having a porous structure wherein a functional group for attachment of the spacer has been introduced.
Claim 9
Dependent Claim
24 words
A modified membrane of claim 6, wherein the polymeric material comprises polystyrenes, polysulfones containing aromatic residues, polyesters, polyamides, polycarbonates, polyvinylidene difluoride and polyvinyl acetate.
Claim 10
Dependent Claim
27 words
A modified membrane of claim 2, wherein the membrane is a flat, permeable, polymeric material having a porous structure onto which are grafted moieties containing functional groups.
Claim 11
Dependent Claim
18 words
A modified membrane of claim 8, wherein the polymeric material comprises polysulfones, polytetrafluoroethylene, polyethylene, polypropylene or polyvinyllidene difluoride.
Claim 12
Dependent Claim
36 words
A modified membrane of claim 2, wherein the membrane is a flat, permeable, polymeric material having a porous structure onto which is coated a second polymeric material containing free functional groups for attachment of the spacer.
Claim 13
Dependent Claim
18 words
A modified membrane of claim 10, wherein the polymeric material comprises polysulfones, polytetrafluoroethylene, polyethylene, polypropylene or polyvinylidene difluoride.
Claim 14
Dependent Claim
30 words
A modified membrane of claim 10, wherein the second polymeric material comprises acrylic or methacrylic acid esters having a free alcohol or ester function for attachment of the spacer group.
Claim 15
Dependent Claim
18 words
A modified membrane of claim 2 wherein N is --(CH 2 ) n --, where n is 1-20.
Claim 16
Dependent Claim
22 words
A modified membrane of claim 2 wherein N is --NH--(CH 2 ) m --NHCO--(CH 2 ) m --CO--, where m is 1-6.
Claim 17
Dependent Claim
10 words
A modified membrane of claim 2 wherein N is oligoglycine.
Claim 18
Dependent Claim
19 words
A modified membrane of claim 2 wherein Y and Z are individually selected from the group consisting of ##STR5##
Claim 19
Dependent Claim
47 words
A modified membrane of claim 2 wherein S W represents a nucleoside of the formula: ##STR6## wherein B W' is a nucleoside base having a protecting group for protection of exocyclic amino groups; and W" is a H or a protecting group for the 2' hydroxy group.
Claim 20
Dependent Claim
19 words
A modified membrane of claim 17 wherein W" is a trityl group, an acyl group or a silyl group.
Claim 21
Dependent Claim
63 words
A modified membrane of claim 2 wherein S W represents an amino acid of the formula: ##STR7## wherein U represents an amino acid side chain; W' represents protecting groups for the side chain of the amino acid; W" represents a protecting group for the amino group of the amino acid; W"' represents a protecting group for the carboxy group of the amino acid.
Claim 22
Dependent Claim
19 words
A modified membrane of claim 19 wherein S W is norleucine attached to the linker through its carboxyl group.
Claim 23
Dependent Claim
18 words
A modified membrane of claim 20, wherein the primary amino group of the nucleocine is protected with fluoroenyl-methyloxycarbonyl.
Claim 24
Dependent Claim
17 words
A method of synthesizing an oligonucleotide comprising sequentially coupling nucleotide to a modified membrane of claim 17.
Claim 25
Dependent Claim
18 words
A method of synthesizing a peptide comprising sequentially coupling amino acids to a modified membrane of claim 19.
Claim 26
Independent Claim
151 words
A method of synthesizing an oligonucleotide comprising the steps of: a. providing a modified membrane represented by the formula: P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and S W is a protected nucleoside, S W being bound to the linker through the functional group Z of the linker; b. coupling a nucleoside phosphoramidite to the nucleoside S W to produce a membrane-bound nucleoside-nucleotide having a phosphite triester linkage; c. oxidizing the phosphite triester to form a phosphate triester linkage; and d. sequentially coupling additional nucleoside phosphoramidites to the membrane-bound nucleoside-nucleotide, and after each coupling step, oxidizing the resulting phosphite triester linkage to a phosphate triester to produce a membrane-bound polynucleotide.
Claim 27
Independent Claim
151 words
A method of synthesizing an oligonucleotide comprising the steps of: a. providing a modified membrane represented by the formula: P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and S W is a protected nucleoside, S W being bound to the linker through the functional group Z of the linker; b. coupling a nucleoside phosphoramidite to the nucleoside S W to produce a membrane-bound nucleoside-nucleotide having a phosphite triester linkage; c. oxidizing the phosphite triester to form a phosphate triester linkage; and d. sequentially coupling additional nucleoside phosphoramidites to the membrane-bound nucleoside-nucleotide, and after each coupling step, oxidizing the resulting phosphite triester linkage to a phosphate triester to produce a membrane-bound polynucleotide.
Claim 28
Dependent Claim
14 words
A method of claim 24, wherein the nucleside phosphoramidite is a nucleoside beta-cyanoethyl phosphoramidite.
Claim 29
Dependent Claim
16 words
A method of claim 24, further comprising removing the protecting groups from the membrane bound polynucleotide.
Claim 30
Dependent Claim
14 words
A method of claim 26, wherein the synthesized polynucleotide is cleaved from the membrane.
Claim 31
Dependent Claim
10 words
A membrane-bound polynucleotide produced by the method of claim 24.
Claim 32
Independent Claim
102 words
A method of synthesizing a peptide, comprising the steps of: a. providing a modified membrane of the formula P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or differential functional groups, the linker being bound to the membrane through the functional group X; and S W is a amino acid, S W being bound to the linker through the functional group Z of the linker; and b. sequentially coupling amino acids to S W to produce a membrane-bound peptide.
Claim 33
Independent Claim
102 words
A method of synthesizing a peptide, comprising the steps of: a. providing a modified membrane of the formula P--X--Y--N--Z--S.sup.W wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or differential functional groups, the linker being bound to the membrane through the functional group X; and S W is a amino acid, S W being bound to the linker through the functional group Z of the linker; and b. sequentially coupling amino acids to S W to produce a membrane-bound peptide.
Claim 34
Dependent Claim
18 words
A method of claim 29, further comprising the step of removing the protecting groups from the membrane-bound peptide.
Claim 35
Dependent Claim
14 words
A method of claim 29, wherein the synthesized peptide is cleaved from the membrane.
Claim 36
Dependent Claim
10 words
A membrane-bound peptide produced by the method of claim 29.
Claim 37
Dependent Claim
11 words
A method of claim 29, wherein S W represents a norleucine.
Claim 38
Dependent Claim
30 words
A method of claim 29, wherein a second linker group is attached to S W , the linker group providing an functional group for coupling of subsequent protected amino acids.
Claim 39
Dependent Claim
11 words
A method of claim 34, wherein the linker is p-hydroxymethylphenoxyacetic acid.
Claim 40
Dependent Claim
12 words
A membrane having an oligonucleotide attached through a terminus of the oligonucleotide.
Claim 41
Dependent Claim
12 words
A membrane having an oligonucleotide attached through a terminus of the oligonucleotide.
Claim 42
Dependent Claim
12 words
A membrane having an peptide attached through a terminus of the peptide.
Claim 43
Dependent Claim
12 words
A membrane having an peptide attached through a terminus of the peptide.
Claim 44
Dependent Claim
111 words
A membrane having an attached oligonucleotide or peptide represented by the formula: P--X--Y--N--Z--(--S.sup.W).sub.n wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and (S W ) n represents an oligonucleotide or peptide comprised of protected or partly protected nucleotides or amino acids, wherein n is the number of nucleotide or amino acid units of the oligonucleotide, (S W ) n being bound to the linker through the functional group Z of the linker.
Claim 45
Dependent Claim
111 words
A membrane having an attached oligonucleotide or peptide represented by the formula: P--X--Y--N--Z--(--S.sup.W).sub.n wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and (S W ) n represents an oligonucleotide or peptide comprised of protected or partly protected nucleotides or amino acids, wherein n is the number of nucleotide or amino acid units of the oligonucleotide, (S W ) n being bound to the linker through the functional group Z of the linker.
Claim 46
Dependent Claim
99 words
A membrane having an attached oligonucleotide or peptide represented by the formula: P--X--Y--N--Z--(--S).sub.n wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and (S) n represents an oligonucleotide or peptide, wherein n is the number of nucleotide or amino acid units of the oligonucleotide or peptide, (S) n being bound to the linker through the functional group Z of the linker.
Claim 47
Dependent Claim
99 words
A membrane having an attached oligonucleotide or peptide represented by the formula: P--X--Y--N--Z--(--S).sub.n wherein P is a polymeric membrane; X is a functional group on the membrane; Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X; and (S) n represents an oligonucleotide or peptide, wherein n is the number of nucleotide or amino acid units of the oligonucleotide or peptide, (S) n being bound to the linker through the functional group Z of the linker.
Claim 48
Dependent Claim
41 words
A modified membrane of the formula: ##STR8## wherein P is a polyvinylidene difluoride; m is 1 through 6; B W is a nucleoside base having a protecting group for protection of exocylic amino groups; and DMT is the 4,4-dimethoxytrityl protecting group.
Claim 49
Dependent Claim
41 words
A modified membrane of the formula: ##STR8## wherein P is a polyvinylidene difluoride; m is 1 through 6; B W is a nucleoside base having a protecting group for protection of exocylic amino groups; and DMT is the 4,4-dimethoxytrityl protecting group.
Claim 50
Dependent Claim
36 words
A modified membrane of the formula: ##STR9## wherein P is a membrane comprising polypropylene coated with hydroxyporpylacrylate; n=1-20; Nle is norleucine; U W is an amino acid side chain; and Fmoc is a fluorenylmethoxycarbonyl protecting group.
Claim 51
Dependent Claim
36 words
A modified membrane of the formula: ##STR9## wherein P is a membrane comprising polypropylene coated with hydroxyporpylacrylate; n=1-20; Nle is norleucine; U W is an amino acid side chain; and Fmoc is a fluorenylmethoxycarbonyl protecting group.
Claim 52
Dependent Claim
22 words
A modified membrane of claim 33, wherein an amino acid is linked to norleucine by ester linkage with a benzyl alcoholic linker.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-05-07 | 1993-11-06 | 1994-05-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-05-07 | 1997-11-06 | 1998-05-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-05-07 | 2001-11-06 | 2002-05-08 | $7700.00 | Window Closed / Lapsed |