US5059654A — Affinity matrices of modified polysaccharide supp…

Claim 1

Independent Claim 76 words

A modified polysaccharide material which comprises: (1) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer made from (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group.

Claim 2

Dependent Claim 76 words

A modified polysaccharide material which comprises: (1) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer made from (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group.

Claim 3

Dependent Claim 12 words

The material of claim 1 wherein said synthetic polymer is a homopolymer.

Claim 4

Dependent Claim 18 words

The material of claim 2 wherein said synthetic polymer is a homopolymer of glycidyl acrylate or glycidyl methacrylate.

Claim 5

Dependent Claim 12 words

The material of claim 1 wherein said synthetic polymer is a copolymer.

Claim 6

Dependent Claim 10 words

The material of claim 1 wherein said polysaccharide is cellulose.

Claim 7

Dependent Claim 22 words

The material of claim 1 wherein said chemical group capable of causing said covalent coupling has been reacted with an affinity ligand.

Claim 8

Dependent Claim 36 words

The material of claim I wherein said chemical group capable of causing the covalent coupling of said synthetic polymer to said affinity ligand is selected from the group consisting of a primary amine and an aldehyde.

Claim 9

Dependent Claim 33 words

The material of claim 6 wherein said affinity ligand is an enzyme, a nucleic acid, an antigen, an antibody, a saccharide, a lectin, an enzyme cofactor, an enzyme inhibitor or a binding protein.

Claim 10

Dependent Claim 24 words

The material of claim 8 wherein said ligand is selected from the group consisting of benzamidine, Protein A, Protein G, polymyxin-B, protamine and heparin.

Claim 11

Dependent Claim 12 words

A self-supporting cellulosic fibrous matrix which comprises the material of claim 6.

Claim 12

Dependent Claim 20 words

In a method of affinity chromatography, the improvement comprising utilizing as the insoluble ligand support the material of claim 1.

Claim 13

Dependent Claim 29 words

In a method of carrying out chemical reactions using an insolubilized biologically active molecule, the improvement wherein the insoluble support for said molecule is the material of claim 1.

Claim 14

Dependent Claim 23 words

The methods of any one of claims 11 or 12 wherein said material is in the form of a self-supporting cellulosic fibrous matrix.

Claim 15

Dependent Claim 92 words

A process for preparing the modified polysaccharide material of claim 1 which comprises: (1) polymerizing said compound (a) which has an epoxy group capable of reacting with the hydroxy group of said polysaccharide, with said compound (b) in the presence of said polysaccharide, under temperature conditions insufficient to cause the covalent binding of said compound (a) to said polysaccharide, to thereby form a synthetic polymer of (a) and (b); (2) reacting said polysaccharide with said chemical group of compound (a) of said synthetic polymer under conditions sufficient to cause said covalent bonding.

Claim 16

Dependent Claim 73 words

A chromatography column for effecting chromatographic separation of at least two components of a sample flowing therethrough comprising: a housing; at least one solid stationary phase in said housing, comprising a matrix having chromatographic functionality and being effective for chromatographic means for distributing the sample through the stationary phase; and means for collecting the sample after the sample has flowed through the stationary phase, wherein said matrix comprises the material of claim 1.

Claim 17

Dependent Claim 115 words

A chromatography column for effecting chromatographic separation of at least two components of a sample flowing therethrough comprising: (1) a housing, said housing comprising: (a) an inlet housing member, and (b) an outlet housing member, said inlet housing member and said outlet housing member defining a radially, outwardly expanding stationary phase chamber; and (2) a stationary phase within said radially outwardly expanding stationary phase chamber, said stationary phase chamber comprising a matrix having chromatographic functionality and being effective for chromatographic separation; wherein said stationary phase and said radially outwardly expanding stationary phase chamber coact to provide substantially uniform radial distribution of sample across said stationary phase, wherein said matrix comprises the material of claim 1.

Claim 18

Dependent Claim 99 words

A chromatography column for effecting chromatographic separation of at least two components of a sample flowing therethrough comprising: (1) a housing, said housing comprising: (a) an inlet housing member, and (b) an outlet housing member, said inlet housing member and said outlet housing member defining a stationary phase chamber; and (2) a stationary phase within said stationary phase chamber, said stationary phase comprising a matrix having chromatographic functionality and being effective for chromatographic separation, wherein said stationary phase chamber and said stationary phase coact to distribute sample across said stationary phase, said matrix comprising the material of claim 1.

Claim 19

Dependent Claim 103 words

A chromatography column for effecting chromatographic separation of at least two components of a sample flowing therethrough comprising: (1) a housing, said housing comprising: (a) an inlet housing member, and (b) an outlet housing member, said inlet housing member and said outlet housing member defining a radially, outwardly expanding stationary phase chamber; and (2) a stationary phase within said radially, outwardly expanding chamber, said stationary phase comprising: (a) matrix having chromatographic functionality and being effective for chromatographic separation, wherein said stationary phase chamber and said stationary phase coact to distribute sample across said stationary phase, said matrix comprising the material of claim 1.

Claim 20

Dependent Claim 103 words

A process for effecting chromatographic separation of at least two components of a sample comprising contacting said sample with a self-supporting fibrous matrix comprising a modified polysaccharide material, said modified polysaccharide material comprising: (1) a water insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer comprising (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group.

Claim 21

Dependent Claim 103 words

A process for effecting chromatographic separation of at least two components of a sample comprising contacting said sample with a self-supporting fibrous matrix comprising a modified polysaccharide material, said modified polysaccharide material comprising: (1) a water insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer comprising (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group.

Claim 22

Dependent Claim 17 words

The process of claim 19, wherein said contacting comprises radial flow of said sample through said matrix.

Claim 23

Dependent Claim 17 words

The process of claim 19, wherein said contacting comprises tangential flow of said sample across said matrix.

Claim 24

Dependent Claim 108 words

A chromatographic device for effecting chromatographic separation of at least two components of a sample, comprising: (A) a modified polysaccharide material which comprises: (1) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer made from (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group; and (B) means for effecting tangential flow of said sample across said modified polysaccharide material.

Claim 25

Dependent Claim 108 words

A chromatographic device for effecting chromatographic separation of at least two components of a sample, comprising: (A) a modified polysaccharide material which comprises: (1) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (2) said synthetic polymer made from (a) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (b) one or more polymerizable compounds containing (i) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (ii) a hydrophobic chemical group; and (B) means for effecting tangential flow of said sample across said modified polysaccharide material.

Claim 26

Dependent Claim 22 words

The device of claim 22, wherein said modified polysaccharide material is in the form of a sheet, a corrugated sheet, or tube.

Claim 27

Dependent Claim 168 words

A chromatography device for effecting chromatographic separation of at least two components of a sample, comprising: (1) a cylindrical core, (2) at least a first and second stationary phases wound around said cylindrical core, wherein (a) a first phase comprises a modified polysaccharide material which comprises: (i) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (ii) said synthetic polymer made from (1) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (2) one or more polymerizable compounds containing (a) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (b) a hydrophobic chemical group, wherein said first phase is in the form of a sheet; (b) a second phase comprising a means for supporting, separating, and providing channels tangential to said cylindrical core; (3) means for distributing the sample to said channels; (4) means for collecting the sample from said channels; and (5) a cylindrical housing.

Claim 28

Dependent Claim 168 words

A chromatography device for effecting chromatographic separation of at least two components of a sample, comprising: (1) a cylindrical core, (2) at least a first and second stationary phases wound around said cylindrical core, wherein (a) a first phase comprises a modified polysaccharide material which comprises: (i) a water-insoluble polysaccharide covalently bonded to a synthetic polymer; (ii) said synthetic polymer made from (1) a polymerizable compound which has an epoxy group capable of direct covalent coupling to said polysaccharide; and (2) one or more polymerizable compounds containing (a) a chemical group capable of causing the covalent coupling of said synthetic polymer to an affinity ligand or a biologically active molecule, or (b) a hydrophobic chemical group, wherein said first phase is in the form of a sheet; (b) a second phase comprising a means for supporting, separating, and providing channels tangential to said cylindrical core; (3) means for distributing the sample to said channels; (4) means for collecting the sample from said channels; and (5) a cylindrical housing.

Claim 29

Dependent Claim 51 words

The device of claim 24, further comprising a nozzle disposed on the end of said cylindrical housing, wherein said means for collecting the sample from said channels comprises one or more grooves along the axial length of said cylindrical core which are in fluid communication with said channels and said nozzle.

Claim 30

Dependent Claim 51 words

The device of claim 24, further comprising a nozzle disposed on the end of said cylindrical housing, wherein said means for distributing the sample to said channels comprises one or more grooves along the axial length of said cylindrical housing which are in fluid communication with said channels and said nozzle.

Claim 31

Dependent Claim 60 words

The device of claim 24, wherein said means for separating, spacing, and providing channels comprises two arrays of filaments, comprising (1) a first array spaced parallel to one another and perpendicular to the axis of said cylindrical core, and (2) a second array of filaments spaced parallel to one another and disposed at an angle to to said first array.

Claim 32

Dependent Claim 19 words

The device of claim 27, wherein said first array and said second array are affixed at their crossover points.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1994-10-21	1995-04-22	1995-10-22	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1998-10-21	1999-04-22	1999-10-22	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2002-10-21	2003-04-22	2003-10-22	$7700.00	Window Closed / Lapsed

Affinity matrices of modified polysaccharide supports

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