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US4411832A
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Polysaccharide matrices comprising macromolecular spacer arms for use as adsorbents in affinity chromatography techniques
Assignee: Individual
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Filing: 1981-07-27
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Granted: 1983-10-25
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Expires: 2001-07-27
Abstract
Abstract Polysaccharide matrices, including polyfunctional macromolecular spacer "arms" coupled to the backbones of such matrices in multipoint attachment, are useful as adsorbents in affinity chromatography techniques to "separate" the matrix from the specific liquid being chromatographed. Exemplary preferred polyfunctional macromolecules include [1] poly-L-lysine, [2] the graft copolymer, poly(L-lysyl-DL-alanine), [3] native albumin and [4] denatured albumin.
Inventors
- 👤 Pedro Cuatrecasas
- 👤 Indu Parikh
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 13:54
CPC Classifications
B; B01; B01J; B01J20/00; B01J20/30; B01J20/32; B01J20/3214; B01J20/3217; B01J20/3219; B01D; B01D15/00; B01D15/08; B01D15/26; B01D15/38; B01D15/3804; B01J20/281; B01J20/286; B01J20/289; B01J20/3231; B01J20/3242; B01J20/3268; B01J20/3272; C; C07; C07J; C07J41/00; C07J41/0033; C07J41/0072
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
85 words
In a polysaccharide matrix useful as an adsorbent for affinity chromatography techniques, the improvement which comprises a plurality of polyfunctional, water-soluble macromolecular spacers individually covalently bonded to the backbone of said polysaccharide matrix in direct multipoint covalent attachment thereto, the said polyfunctional macromolecular spacers being selected from the group consisting of (1) poly-L-lysine, (2) the graft copolymer, poly(L-lysyl-DL-alanine), (3) native albumin and (4) denatured albumin and the endstanding functional groups of said polyfunctional macromolecular spacers being essentially sterically unhindered with respect to said polysaccharide matrix.
Claim 2
Dependent Claim
85 words
In a polysaccharide matrix useful as an adsorbent for affinity chromatography techniques, the improvement which comprises a plurality of polyfunctional, water-soluble macromolecular spacers individually covalently bonded to the backbone of said polysaccharide matrix in direct multipoint covalent attachment thereto, the said polyfunctional macromolecular spacers being selected from the group consisting of (1) poly-L-lysine, (2) the graft copolymer, poly(L-lysyl-DL-alanine), (3) native albumin and (4) denatured albumin and the endstanding functional groups of said polyfunctional macromolecular spacers being essentially sterically unhindered with respect to said polysaccharide matrix.
Claim 3
Dependent Claim
24 words
The polysaccharide matrix as defined by claim 1, wherein the polyfunctional macromolecule is poly(L-lysyl-DL-alanine), a graft copolymer of poly-L-lysine (backbone) and poly-DL-alanine (side chains).
Claim 4
Dependent Claim
28 words
The polysaccharide matrix as defined by claim 2, wherein the poly(L-lysyl-DL-alanine) has an average molecular weight of about 37,500 and a lysine to alanine ratio of about 1:15.
Claim 5
Dependent Claim
14 words
The polysaccharide matrix as defined by claim 1, wherein the polyfunctional macromolecule is poly-L-lysine.
Claim 6
Dependent Claim
19 words
The polysaccharide matrix as defined by claim 4, wherein the poly-L-lysine has an average molecular weight of about 160,000.
Claim 7
Dependent Claim
15 words
The polysaccharide matrix as defined by claim 1, wherein the polyfunctional macromolecule is native albumin.
Claim 8
Dependent Claim
15 words
The polysaccharide matrix as defined in claim 6, wherein the native albumin is bovine albumin.
Claim 9
Dependent Claim
15 words
The polysaccharide matrix as defined by claim 1, wherein the polyfunctional macromolecule is denatured albumin.
Claim 10
Dependent Claim
16 words
The polysaccharide matrix as defined by claim 8, wherein the denatured albumin is urea-denatured bovine albumin.
Claim 11
Dependent Claim
24 words
The polysaccharide matrix as defined by claim 1, wherein the polysaccharide is selected from the group consisting of cellulose, starch, cross-linked dextran, and agarose.
Claim 12
Dependent Claim
21 words
The polysaccharide matrix as defined by claim 1, wherein the said polysaccharide-polyfunctional macromolecular complex is diluted with an unsubstituted polysaccharide gel.
Claim 13
Dependent Claim
17 words
The polysaccharide matrix as defined by claim 11, wherein the said dilution is about 200- to 600-fold.
Claim 14
Dependent Claim
39 words
In a method for the affinity chromatography by technique of coupling ligands, proteins, hormones, nucleotides and nucleosides to a polysaccharide matrix, the improvement which comprises coupling said biologically active molecules to the polysaccharide matrix as defined by claim 1.
Claim 15
Dependent Claim
34 words
The method as defined by claim 13, wherein the molecule coupled to the said polysaccharide matrix is selected from the group consisting of ligands bearing an amino function and ligands bearing a carboxyl function.
Claim 16
Dependent Claim
25 words
The method as defined by claim 14, wherein the ligand is separated from the polysaccharide matrix by a distance in the order of 150 Å.
Claim 17
Dependent Claim
10 words
The product of the method as defined by claim 13.
Claim 18
Dependent Claim
10 words
The product of the method as defined by claim 14.
Claim 19
Dependent Claim
27 words
In an affinity chromatography column comprising an insoluble, solid polysaccharide adsorbent, the improvement which comprises, as the adsorbent thereof, the polysaccharide matrix as defined by claim 1.
Claim 20
Dependent Claim
65 words
In a polysaccharide matrix useful as an adsorbent for affinity chromatography techniques, the improvement which comprises a plurality of polyfunctional, water-soluble macromolecule spacers individually covalently bonded to the backbone of said polysaccharide matrix in direct multipoint covalent attachment thereto, the polyfunctional macromolecule comprising endstanding functional groups, and the endstanding functional groups of said polyfunctional macromolecule being essentially sterically unhindered with respect to said polysaccharide matrix.
Claim 21
Dependent Claim
65 words
In a polysaccharide matrix useful as an adsorbent for affinity chromatography techniques, the improvement which comprises a plurality of polyfunctional, water-soluble macromolecule spacers individually covalently bonded to the backbone of said polysaccharide matrix in direct multipoint covalent attachment thereto, the polyfunctional macromolecule comprising endstanding functional groups, and the endstanding functional groups of said polyfunctional macromolecule being essentially sterically unhindered with respect to said polysaccharide matrix.
Claim 22
Dependent Claim
16 words
The polysaccharide matrix as defined by claim 19, wherein the polyfunctional macromolecule is a polyamino macromolecule.
Claim 23
Dependent Claim
21 words
The polysaccharide matrix as defined by claim 19, wherein the endstanding functional groups of said polyfunctional macromolecule comprise a succinyl moiety.
Claim 24
Dependent Claim
21 words
The polysaccharide matrix as defined by claim 19, wherein the endstanding functional groups of said polyfunctional macromolecule comprise a hydrazide moiety.
Claim 25
Dependent Claim
21 words
The polysaccharide matrix as defined by claim 19, wherein the endstanding functional groups of said polyfunctional macromolecule comprises a p-aminophenyl moiety.
Claim 26
Dependent Claim
20 words
The polysaccharide matrix as defined by claim 19, wherein the endstanding functional groups of said polyfunctional macromolecule comprise a ##STR2##
Claim 27
Dependent Claim
38 words
The polysaccharide matrix as defined by claim 19, wherein the endstanding functional groups of said polyfunctional macromolecule comprise a moiety selected from the group consisting of α of ξ NH 2 , carboxyl, azide, ester and diazonium salt.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-10-24 | 1987-04-25 | 1987-10-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-10-24 | 1991-04-25 | 1991-10-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-10-24 | 1995-04-25 | 1995-10-25 | $7700.00 | Window Closed / Lapsed |