Patent US3873514A successfully refreshed from Google Patents.
US3873514A
Expired / Lapsed
US
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Preparation of gel for affinity chromatography
Assignee: Bio Rad Laboratories Inc
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Filing: 1974-05-17
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Granted: 1975-03-25
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Expires: 1994-05-17
Abstract
Abstract Improved functionalized agarose beads are provided which find use in affinity chromatography. The agarose beads are first cross-linked with a polyfunctional compound under mild conditions, followed by reacting with a haloalkylcarboxy compound or, alternatively, a haloalkylamine compound followed by amide formation with a cyclic carboxylic acid anhydride, to provide a final product having an ether linkage and a carboxy functionality for linking to other compounds. The resulting agarose beads have improved flow and packing characteristics and improved chemical and physical stability, while providing a stable functionality for linking to a wide variety of compounds.
Inventors
- 👤 Albert E Y Chu
- 👤 Christopher J Siebert
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 19:23
CPC Classifications
C; C08; C08B; C08B37/00; C08B37/0006; C08B37/0036; C08B37/0039; B; B01; B01J; B01J20/00; B01J20/281; B01J20/286; B01J20/291
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
83 words
A METHOD FOR IMPROVING THE PHYSICAL AND CHEMICAL CHARACTERISTICS OF AGAROSE BEADS WHILE OBTAINING IMPROVED FLOW AND PACKING PROPERTIES COMPRISING: A. COMBINING IN AN AQUEOUS MEDIUM UNDER MILD CONDITIONS AT AN ALKALINE PH AGAROSE BEADS WITH 1,3-DIHALOPROPANOL2, WHEREIN THE HALOGEN IS OF ATOMIC NUMBER 17 TO 35, IN AN AMOUNT OF FROM ABOUT 0.5 TO 50 MMOLE PER GRAM OF AGAROSE, FOR A TIME SUFFICIENT TO PROVIDE THE DESIRED DEGREE OF CROSS-LINKING; AND B. WASHING THE CROSS-LINKED AGAROSE BEADS FREE OF UNREACTED 1,3-DIHALOPROPANOL-2.
Claim 2
Independent Claim
83 words
A METHOD FOR IMPROVING THE PHYSICAL AND CHEMICAL CHARACTERISTICS OF AGAROSE BEADS WHILE OBTAINING IMPROVED FLOW AND PACKING PROPERTIES COMPRISING: A. COMBINING IN AN AQUEOUS MEDIUM UNDER MILD CONDITIONS AT AN ALKALINE PH AGAROSE BEADS WITH 1,3-DIHALOPROPANOL2, WHEREIN THE HALOGEN IS OF ATOMIC NUMBER 17 TO 35, IN AN AMOUNT OF FROM ABOUT 0.5 TO 50 MMOLE PER GRAM OF AGAROSE, FOR A TIME SUFFICIENT TO PROVIDE THE DESIRED DEGREE OF CROSS-LINKING; AND B. WASHING THE CROSS-LINKED AGAROSE BEADS FREE OF UNREACTED 1,3-DIHALOPROPANOL-2.
Claim 3
Dependent Claim
52 words
A method according to claim 1, wherein said halo is chloro and is present in from about 1 to 25 mmole per gram of agarose, said beads are present in said aqueous medium in from about 0.5 to 10 weight percent and said pH is in the range of 9 to 13.
Claim 4
Dependent Claim
83 words
A method according to claim 1, including the additional steps of: c. combining said cross-linked agarose beads in an aqueous medium at an alkaline pH under mild conditions with a compound of the formula: X-R-N(H)Y wherein: X is halogen of atomic number 17 to 35; R is alkylene of from 2 to 6 carbon atoms; Y is hydrogen or is an acyl residue of a dicarboxylic acid capable of forming a cyclic anhydride of from 5 to 6 annular members of the formula:
Claim 5
Dependent Claim
10 words
A method according to claim 3, wherein Y is hydrogen.
Claim 6
Dependent Claim
12 words
A method according to claIm 3, wherein Y is said acryl residue.
Claim 7
Dependent Claim
55 words
A method according to claim 3, including the addtional steps of: d. isolating said agarose beads; and e. repeatedly washing said agarose beads with an inert substantially anhydrous polar organic solvent, followed by separation of said agarose beads from said solvent, until the water content of said agarose beads is below about 2 weight percent.
Claim 8
Dependent Claim
26 words
A method according to claim 6, wherein said organic solvent is at least one of an alkanol of from 1 to 4 carbon atoms or dioxane.
Claim 9
Dependent Claim
47 words
A method according to claim 6, including the additional step of: f. combining said dehydrated agarose beads in an inert polar solvent with an at least about a stoichiometric amount based on carboxylic acid groups of N-hydroxy succinimide and carbodiimide, to form the ester of N-hydroxy succinimide.
Claim 10
Dependent Claim
36 words
Agarose beads of from about 15 to 500 mesh, having exclusion limits of from about 1 to 50 X 106 Daltons and having from about 0.0005 to 0.5 moles of 2-hydroxy-1,3-propylene cross-linking per gram of agarose.
Claim 11
Dependent Claim
36 words
Agarose beads of from about 15 to 500 mesh, having exclusion limits of from about 1 to 50 X 106 Daltons and having from about 0.0005 to 0.5 moles of 2-hydroxy-1,3-propylene cross-linking per gram of agarose.
Claim 12
Dependent Claim
64 words
Agarose beads according to claim 9, having from about 0.0005 to 1 mole of: -R-N(H)Y groups bound to said agarose through an ether linkage; wherein: R is alkylene of from 2 to 6 carbon atoms; and Y is hydrogen or is an acyl residue of a dicarboxylic acid capable of forming a cyclic anhydride of from 5 to 6 annular members of the formula:
Claim 13
Dependent Claim
7 words
Agarose beads according to claim 10, wherein
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) | 1978-03-24 | 1978-09-23 | 1979-03-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-03-24 | 1982-09-23 | 1983-03-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-03-24 | 1986-09-23 | 1987-03-25 | $7700.00 | Window Closed / Lapsed |