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US3960720A Expired / Lapsed US ⏱️ 0.0 years remaining

Gel product for separation purposes and method of using the product for hydrophobic salting out adsorption

Assignee: Exploaterings TBF AB
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Filing: 1974-03-19
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Granted: 1976-06-01
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Expires: 1994-03-19
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Abstract

Abstract The invention concerns a gel product for chromatography based on agarose, and a method of using the product for hydrophobe salting out adsorption. The gel product is characterized in that the hydrophobe groups are bonded to the agarose with ether or esther binding, and that the hydrophobe groups can be composed of aliphatic hydrocarbons, halogen or hydroxy-substituted alkyl groups, with 1 to 20 carbon atoms, or contain 1 or more, up to 5, aromatic nuclei, unsubstituted or substituted with alkyl, alkyloxy, halogen or NO2 or have the formula OH ¦ -CH2-CH-CH2-O-R where R can be alkyl, alkenyl, aryl, aralkyl, aralkoxy, alkaryl, optionally substituted with hydroxy, halogen or nitro, and where the alkyl parts of R contain 1 to 20 carbon atoms. The method of separating the amphipatic substances is based on the fact that they are adsorbed by an amphipatic gel in which the salt concentration in the solution is adjusted beyond the level at which the ion concentration least corresponds to the ion concentration in an aqeous solution of 1 M NaCl, and desorption takes place through lowering the ion concentration and/or lowering the pH and/or lowering the polarity of the solvent. The amphipatic substances are primarily proteins or peptides.

Inventors

  • 👤 Jerker Olof Porath
  • 👤 Jan Gunnar Rosengren

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-24 14:18
CPC Classifications
B; B01; B01J; B01J20/00; B01J20/22; C; C08; C08B; C08B37/00; C08B37/0006; C08B37/0036; C08B37/0039; B01D; B01D15/00; B01D15/08; B01D15/26; B01D15/32; B01D15/325; B01D15/327

Claims Analysis

Total Claims: 6
Claim 1
Independent Claim 267 words
Method of separating proteins containing both hydrophilic groups such as OH, NH 2 , COOH and hydrophobic groups such as alkyl, phenyl, indyl, comprising adsorbing the proteins from a solvent onto a gel packed in a column and consisting essentially of agarose to which neutral hydrophobic groups which do not form ions are covalently bonded and having the formula P--O(CO).sub.n.sub.' --R where P is the agarose residue and R is a hydrophobic radical selected from the group consisting of hydrocarbon and ether residues, n' being no greater than 1, the hydrophobic group having the formula C n H 2n +1 unsubstituted or substituted with halogen or hydroxy, C n H 2n-1 , X m C 6 H 5-m --, X m C 6 H 5-m CY--, X m C 6 H 5-m CY--CZ--, X m C 10 H 7-m --, X m C 10 H 7-m CY--, X m C 10 H 7-m CY--CZ-- or ##EQU3## where X is H, C p H 2p +1 , CH 3 , C 2 H 5 , OC p H 2p +1 , halogen or NO 2 , n is an integer from 1 to 20, Y and Z are O or H 2 , m is an integer from 0 to 5, p is an integer from 1 to 10 and R' is alkyl, alkenyl, aryl, aralkyl, aralkoxy, alkaryl, unsubstituted or substituted with hydroxy, halogen or nitro, the alkyl portions of R' containing 1- 20 carbon atoms, and desorbing the adsorbed material by lowering of the ion strength and/or changing the pH and/or lowering the polarity of the solvent.
Claim 2
Dependent Claim 267 words
Method of separating proteins containing both hydrophilic groups such as OH, NH 2 , COOH and hydrophobic groups such as alkyl, phenyl, indyl, comprising adsorbing the proteins from a solvent onto a gel packed in a column and consisting essentially of agarose to which neutral hydrophobic groups which do not form ions are covalently bonded and having the formula P--O(CO).sub.n.sub.' --R where P is the agarose residue and R is a hydrophobic radical selected from the group consisting of hydrocarbon and ether residues, n' being no greater than 1, the hydrophobic group having the formula C n H 2n +1 unsubstituted or substituted with halogen or hydroxy, C n H 2n-1 , X m C 6 H 5-m --, X m C 6 H 5-m CY--, X m C 6 H 5-m CY--CZ--, X m C 10 H 7-m --, X m C 10 H 7-m CY--, X m C 10 H 7-m CY--CZ-- or ##EQU3## where X is H, C p H 2p +1 , CH 3 , C 2 H 5 , OC p H 2p +1 , halogen or NO 2 , n is an integer from 1 to 20, Y and Z are O or H 2 , m is an integer from 0 to 5, p is an integer from 1 to 10 and R' is alkyl, alkenyl, aryl, aralkyl, aralkoxy, alkaryl, unsubstituted or substituted with hydroxy, halogen or nitro, the alkyl portions of R' containing 1- 20 carbon atoms, and desorbing the adsorbed material by lowering of the ion strength and/or changing the pH and/or lowering the polarity of the solvent.
Claim 3
Dependent Claim 36 words
Method according to claim 1, and adjusting the salt level of said solution above the level where the ion strength at least corresponds to the ion strength in an aqueous solution of 1 M sodium chloride.
Claim 4
Dependent Claim 220 words
Gel product for hydrophobic salting out chromatographic adsorption for the separation of protein fractions, consisting essentially of agarose to which neutral hydrophobic groups which do not form ions are covalently bonded and having the formula P--O(CO).sub.n .sub.'--R where P is the agarose residue and R is a hydrophobic radical selected from the group consisting of hydrocarbon and ether residues, n' being no greater than 1, the hydrophobic group having the formula C n H 2n +1 unsubstituted or substituted with halogen or hydroxy, C n H 2n-1 , X m C 6 H 5-m --, X m C 6 H 5-m CY--, X m C 6 H 5-m CY-CZ--, X m C 10 H 7-m --, X m C 10 H 7-m CY--, X m C 10 H 7-m CY--CZ-- or ##EQU4## where X is H, C p H 2p +1 , CH 3 , C 2 H 5 , OC p H 2p +1 , halogen or NO 2 , n is an integer from 1 to 20, Y and Z are O or H 2 , m is an integer from 0 to 5, p is an integer from 1 to 10 and R' is alkyl, alkenyl, aryl, aralkyl, aralkoxy, alkaryl, unsubstituted or substituted with hydroxy, halogen or nitro, the alkyl portions of R' containing 1-20 carbon atoms.
Claim 5
Dependent Claim 220 words
Gel product for hydrophobic salting out chromatographic adsorption for the separation of protein fractions, consisting essentially of agarose to which neutral hydrophobic groups which do not form ions are covalently bonded and having the formula P--O(CO).sub.n .sub.'--R where P is the agarose residue and R is a hydrophobic radical selected from the group consisting of hydrocarbon and ether residues, n' being no greater than 1, the hydrophobic group having the formula C n H 2n +1 unsubstituted or substituted with halogen or hydroxy, C n H 2n-1 , X m C 6 H 5-m --, X m C 6 H 5-m CY--, X m C 6 H 5-m CY-CZ--, X m C 10 H 7-m --, X m C 10 H 7-m CY--, X m C 10 H 7-m CY--CZ-- or ##EQU4## where X is H, C p H 2p +1 , CH 3 , C 2 H 5 , OC p H 2p +1 , halogen or NO 2 , n is an integer from 1 to 20, Y and Z are O or H 2 , m is an integer from 0 to 5, p is an integer from 1 to 10 and R' is alkyl, alkenyl, aryl, aralkyl, aralkoxy, alkaryl, unsubstituted or substituted with hydroxy, halogen or nitro, the alkyl portions of R' containing 1-20 carbon atoms.
Claim 6
Dependent Claim 17 words
Gel product according to claim 3, in which the hydrophobic groups are β-hydroxy propyl or β-hydroxy butyl.

References Cited (Backward Citations)

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US3285849A Cited
US3350174A Cited
US3453257A Cited
US3527712A Cited
US3536614A Cited
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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) 1979-06-01 1979-12-01 1980-06-01 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1983-06-01 1983-12-01 1984-06-01 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1987-06-01 1987-12-01 1988-06-01 $7700.00 Window Closed / Lapsed

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