Affinity adsorbents for fibrinogen
For the separation, removal, isolation, purification, characterization, identification or quantification of fibrinogen or a protein that is a fibrinogen analogue, an affinity adsorbent is used that is a compound of formula II wherein one X is N and the other is N, C—Cl or C—CN; Y is O, S or NR 2 ; 0 Z is O, S or NR 3 ; R 2 and R 3 are each H, alkyl, hydroxyalkyl, benzyl or &bgr; -phenylethyl; n is 0 to 6; A is a support matrix, optionally linked to the triazine ring by a spacer; R 7 is a group bearing a positive charge at neutral pH; W is an optional linker; V is an aromatic group; and R 8 and R 9 are each H, OH, alkyl, alkoxy, amino, NH 2 , acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, sulphamoyl, alkylsulphonyl or halogen or a cyclic structure such as a morpholino group, or R 8 and R 9 are linked to form such a cyclic structure.
1. A method for purifying fibrinogen or a protein that is a fibrinogen analogue, wherein said method comprises contacting a sample containing fibrinogen or a protein that is a fibrinogen analogue with an affinity adsorbent of formula II:
wherein 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;
n is 0 to 6;
A is a support matrix selected from agarose, silica, cellulose, dextran, starch, alginate, carrageenan, synthetic polymers, glass and metal oxides, wherein A is optionally linked to the triazine ring by a spacer;
R 7 is a group bearing a positive charge at neutral pH;
W is an optional linker; and
either V is an aromatic group, R 8 is H, OH, alkyl, alkoxy, amino, acyloxy, acylamino, CO 2 H, sulphonic acid, carbamoyl, sulphamoyl, alkylsulphonyl or halogen and R 9 is a morpholino group, or VR 8 R 9 is morpholine,
wherein the fibrinogen, or the protein that is a fibrinogen analogue, binds to the affinity adsorbent; and
wherein said method further comprises recovering the fibrinogen or the protein that is a fibrinogen analogue from the affinity adsorbent, thereby obtaining purified fibrinogen or purified fibrinogen analogue with a purity of greater than 85%.
2. The method, according to claim 1 , wherein the adsorbent is of formula III:
3. The method, according to claim 1 , wherein the adsorbent is of formula IV:
4. The method, according to claim 1 , wherein the adsorbent is of formula V:
5. The method, according to claim 1 , wherein the adsorbent is of formula VI:
6. The method, according to claim 1 , wherein the adsorbent is of formula VII:
7. The method according to claim 1 , wherein the fibrinogen is in a sample of plasma.