Patent US5831321A successfully refreshed from Google Patents.
EP0064833A2
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EP
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High pressure liquid affinity chromatography
Assignee: The AD Centre
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Filing: 1982-04-26
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Granted: —
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Expires: 2002-04-26
Abstract
Abstract A process for the high pressure liquid affinity chroma- tographis separation of at least one biological or related substance from a mixture in which the contact, washing and eluting phases are performed on a binding material made from a ligand. contair'.ig at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm, the binding material being so constructed that at least one biological or related substance is retained on the binding material during the contact and washing phases. In one preferred embodiment the ligand is a reactive dye. especially a triazinyl dye, the matrix is agarose or silica and the spacer arm is a substituted aminohexyl group. The chromatographic procedure is preferably performed at 100-3500 p si, at a flow rate of 0.5-2.0ml/min. The choice of washing and eluting solutions depends on the material to be separated. However buffer solution to wash the column and a desorbing agent to elute the material are preferred. When the biological substance is an enzyme the desorbing agent is preferably an enzyme substrate, cofactor, inhibitor or analogue thereof. A binding material for use in the above HPLAC procedure is also provided. This is made from a ligand containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm.
Inventors
- 👤 Anthony Atkinson
- 👤 Christopher Robin Lowe
- 👤 Klaus University Of Lund Mosbach
- 👤 David Anthony Philip Small
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-09 03:49
CPC Classifications
B; B01; B01D; B01D15/00; B01D15/08; B01D15/26; B01D15/38; B01D15/3804; B01D15/3828; B01J; B01J20/00; B01J20/22; B01J20/26; B01J20/265; B01J20/267; B01J20/281; B01J20/286; B01J20/289; B01J20/30; B01J20/3092; B01J20/32; B01J20/3202; B01J20/3204; B01J20/3206; B01J20/3208; B01J20/321; B01J20/3212; B01J20/3214; B01J20/3217; B01J20/3219; B01J20/3231; B01J20/3242; B01J20/3244; B01J20/3246; B01J20/3257; B01J20/3259; B01J20/3261; B01J20/3263; C; C12; C12N; C12N9/00; B01J2220/00; B01J2220/40; B01J2220/49; B01J2220/50; B01J2220/54; B01J2220/58
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
90 words
A process for the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture wherein contact, washing and elution phases are performed on a binding material characterised in that the binding material comprises a ligand, containing at least one of the groups anthra- .quinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm said binding material being so constructed that the at least one biological or related substance is retained on the binding material during the contact and washing phases.
Claim 2
Dependent Claim
90 words
A process for the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture wherein contact, washing and elution phases are performed on a binding material characterised in that the binding material comprises a ligand, containing at least one of the groups anthra- .quinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm said binding material being so constructed that the at least one biological or related substance is retained on the binding material during the contact and washing phases.
Claim 3
Dependent Claim
81 words
A process according to claim 1 characterised in that the ligand is derived from a compound of general formula III wherein A is an organic substituent containing an anthraquinone, preferably a sulphonated anthraquinone, an aromatic azo or a phthalocyanine group, B is an organic substituent, preferably a sulphonated aromatic group, a halogen atom, preferably a chlorine atom, or an optionally substituted amino group, and C is a leaving group, preferably a chlorine atom, which is displaced in a nucleophilic substitution reaction.
Claim 4
Dependent Claim
32 words
A process according to claim 2 characterised in that the ligand is derived from Cibacron Blue F3G-A, Procion Blue MXR, Procion Green H4G, Procion Yellow HA or Procion Brown MX - 5BR.
Claim 5
Dependent Claim
43 words
A process according to any one of claims 1 to 3 characterised in that the matrix is a cross linked cellulose, cross linked agarose, cross linked dextrose, cross linked dextran, cross linked polyacrylamide, cross linked polyacrylamide-agarose, alumina, titania, zirconia, silica or glass beads.
Claim 6
Dependent Claim
55 words
A process according to any preceding claim characterised in that the binding material has the general formula IV Matrix - X - R - Y - Ligand wherein X-R-Y is the spacer arm and X is an organic or organometallic functional group, R is an organic group and Y is a nucleophilic group or atom.
Claim 7
Dependent Claim
52 words
A process according to claim 5 characterised in that R is one or more of the following groups alkyl, alkene, ∝- aminoalkyl, ∝-.oxyalkyl, ∝-thioalkyl, alkyl alcohol, diol, carboxylate, isourea, carbamate, guanidino, hydrazino, aromatic or heterocyclic and Y is alkyl, ether, thioether, amido, amino, - NH -, - 0 - or - S-.
Claim 8
Dependent Claim
114 words
A process according to either claim 5 or claim 6 characterised in that the binding material has the general formula IV, Matrix - X - R - Y - Ligand IV wherein the matrix is silica, X is wherein R 1 is H or alkyl and R 2 is one or more of the following groups alkyl, substituted alkyl, diol, ether, amido or amino, R is one or more of the following groups alkyl, alkene, ∝-aminoalkyl, ∝-oxyalkyl, ∝-thioalkyl, alkyl alcohol, diol, carboxylate, ether, thioether, amido, amino, amidino, imino carboxylate, isourea, carbamate, guanidino, hydrazino, aromatic or heterocyclic and Y is alkyl, ether, thioether, amido, amino, - NH -, - 0 - or - S -.
Claim 9
Dependent Claim
69 words
A process according to claim 7 characterised in that R 2 is one or more of the following groups alkyl, substituted alkyl or alkyl ether R is (̵ CH 2 )̵ n , - NH(CH 2 )̵ n , -O(CH 2 )̵ n or -S(CH 2 )̵ n , wherein n is from 1 to 12 and Y is - NH -, - 0 - or - S -.
Claim 10
Dependent Claim
59 words
A process according to claim 8 characterised in that the spacer arm (X - R - Y) is -OSi(OR 1 ) 2 (CH 2 ) 3 OCH 2 CH(OH)CH 2 NH(CH 2 ) 6 NH-, -OSi(OR 1 ) 2 CH 2 CH 2 CH 2 S-or -OSi(OR 1 ) 2 (CH 2 ) 3 OCH 2 CH(OH)CH 2 O-.
Claim 11
Dependent Claim
49 words
A process according to any preceding claim characterised in that the at least one biological or related substance is bound to the binding material in the contact phase in the presence of a metal ion selected from the group Co 2+ , Ni 2+ , and Zn 2+ .
Claim 12
Dependent Claim
76 words
A binding material for use in the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture comprising a ligand, containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm said binding material being so constructed that the at least one biological or related substance is retained on the binding material during the contact and washing phases of HPLAC.
Claim 13
Dependent Claim
76 words
A binding material for use in the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture comprising a ligand, containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm said binding material being so constructed that the at least one biological or related substance is retained on the binding material during the contact and washing phases of HPLAC.
Claim 14
Dependent Claim
92 words
A binding material according to claim 11 having the general formula IV Matrix - X - R - Y - Ligand IV wherein the matrix is silica, X is wherein R 1 is H or alkyl and R2 is one or more of the following groups alkyl, substituted alkyl or alkyl ether, R is -(̵CH 2 )̵ n , -NH(CH 2 )̵ n , -O(CH 2 )̵ n or -S(CH 2 )̵ n . wherein n is from 1 to 12, and Y is -NH-, - 0 - or - S -.
References Cited (Backward Citations)
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