Adsorbents for protein purification
View Patent ↗Use of an affinity adsorbent for the separation, removal, isolation, purification, characterisation, identification or quantification of a proteinaceous material, wherein the affinity adsorbent is a compound of formula (III), wherein R 1 is H, alkyl, aryl, hydroxyalkyl, cyciohexyl, amino or a heterocyclic group which is optionally substituted with one or more of alkyl, aryl, alkoxy, aryloxy, acyloxy, acylamino, amino, OH, CO 2 H, sulphonyl, carbamoyl, sulphamoyl, alkylsuiphonyl and halogen; one X is N and the other is N, C—Cl or C—CN; Y is O, S or NR 2 ; Z is O, S or NR 3 ; R 2 and R 3 are each H, alkyl, hydroxyalkyl, benzyl or β-phenylethyl; Q is benzene, naphthalene, benzthiazole, benzoxazole, 1-phenylpyrazole, indazoie or benzimidazoie; R 4 , R 5 and R 6 are each H, OH, alkyl, aryl, heterocyclic, alkoxy, aryloxy, amino, acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, suiphamoyl, alkylsulphonyl or halogen, or two or more of R 4 , R 5 and R 6 are linked to form a cyclic structure; U and V are the same or different C 1-10 straight-chain alkylene groups optionally substituted by one or more of hydroxyl, alkyl, aryl, hydroxyl, alkyl, β-phenylethyl and halogen; and A is a support matrix optionally linked to the X-containing ring by a spacer.
1. A method for purifying a proteinaceous material, wherein said method comprises contacting a sample containing a proteinaceous material with an affinity adsorbent of formula III
wherein R 1 is an aryl or aromatic heterocyclic group which may contain a further fused ring and can be substituted with one or more of alkyl, aryl, alkoxy, aryloxy, acyloxy, acylamino, amino, OH, CO 2 H, sulphonyl, carbamoyl, sulphamoyl, alkylsulphonyl and halogen;
X is N;
Y is O, S or NR 2 ;
Z is O, S or NR 3 ;
R 2 and R 3 are each H, alkyl, hydroxyalkyl, benzyl or β-phenylethyl;
Q is benzene, naphthalene, benzthiazole, benzoxazole, 1-phenylpyrazole, indazole or benzimidazole;
R 4 , R 5 and R 6 are each H, OH, alkyl, aryl, heterocyclic, alkoxy, aryloxy, amino, acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, sulpharnoyl, alkylsulphonyl or halogen, and QR 4 R 5 R 6 is or includes a cyclic structure;
U and V are the same or different C 1-10 straight-chain alkylene groups and U and/or V is substituted by OH; and
A is a support matrix optionally linked to the X-containing ring by a spacer.
2. The method according to claim 1 , wherein either or each cyclic structure has an OH or SO 3 H substituent.
3. The method according to claim 1 , wherein U and/or V is CHOH.
4. A method for purifying a proteinaceous material, wherein said method comprises contacting a sample containing a proteinaceous material with an affinity adsorbent selected from one of the following:
5. The method according to claim 4 , wherein the adsorbent is of formula V
6. The method according to claim 4 , wherein the adsorbent is of formula VI
7. The method according to claim 4 , wherein the adsorbent is of formula VII
8. The method according to claim 1 , wherein the adsorbent is of formula VIII
9. The method according to claim 1 , wherein the adsorbent is of formula IX
10. The method according to claim 1 , wherein the adsorbent is of formula X
11. The method according to claim 1 , wherein the adsorbent is of formula XI
12. The method according to claim 1 , wherein the proteinaceous material is an immunoglobulin, immunoglobulin fragment or protein.
13. The method according to claim 12 , wherein the material is a monoclonal antibody.
14. The method according to claim 12 , wherein the material is an immunoglobulin fragment selected from Fab, Fab′, F(ab′)2, scFV, diabody, minibody, tribody and tetrabody fragments.
15. The method according to claim 1 , wherein the proteinaceous material is in cell culture.
16. The method according to claim 1 , wherein the proteinaceous material is in combination with an anti-foaming agent.
17. The method according to claim 16 , wherein the anti-foaming agent is a block copolymer of polyoxyethylene and polyoxypropylene.
18. A compound of formula III
wherein R 1 is an aryl or aromatic heterocyclic group which may contain a further fused ring and can be substituted with one or more of alkyl, aryl, alkoxy, aryloxy, acyloxy, acylamino, amino, OH, CO 2 H, sulphonyl, carbamoyl, sulphamoyl, alkylsulphonyl and halogen;
X is N;
Y is O, S or NR 2 ;
Z is O, S or NR 3 ;
R 2 and R 3 are each H, alkyl, hydroxyalkyl, benzyl or β-phenylethy;
Q is benzene, naphthalene, benzthiazole, benzoxazole, 1-phenylpyrazole, indazole or benzimidazole;
R 4 , R 5 , and R 6 are each H, OH, alkyl, aryl, heterocyclic, alkoxy, aryloxy, amino, acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, sulphamoyl, alkylsulphonyl or halogen, QR 4 R 5 R 6 is or includes a cyclic structure, and R 1 and/or QR 4 R 5 R 6 has an OH or SO 3 H substituent;
U and V are the same or different C 1-10 straight-chain alkylene groups and U and/or V is substituted by OH and
A is a support matrix optionally linked to the X-containing ring by a spacer.
19. The compound according to claim 18 , wherein U and/or V is CHOH.
20. A method for purifying a proteinaceous material, wherein said method comprises contacting a sample containing a proteinaceous material with an affinity adsorbent of formula III
wherein R 1 is an or aromatic heterocyclic group which may contain a further fused ring and can be substituted with one or more of alkyl, aryl, alkoxy, acyloxy, acyloxy, acylamino, amino, OH, CO 2 H, sulphonyl, carbamoyl, sulphamoyl, alkylsulphonyl and halogen;
X is N;
Y is O, S or NR 2 ;
Z is O, S or NR 3 ;
R 2 and R 3 are each H, alkyl, hydroxyalkyl, benzyl or β-phenylethyl;
Q is benzene, naphthalene, benzthiazole, benzoxazole, 1-phenylpyrazole, indazole or benzimidazole;
R 4 , R 5 and R 6 are each H, OH, alkyl, aryl, heterocyclic, alkoxy, acyloxy, amino, acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, sulphamoyl, alkylsulphonyl or halogen, and QR 4 R 5 R 6 is or includes a cyclic structure;
U and V are the same or different C 1-10 straight-chain alkylene groups and U and/or V is substituted by OH; and
A is a support matrix optionally linked to the X-containing ring by a spacer, and
wherein the proteinaceous material is in combination with an anti-foaming agent.