Use of a chromatography substrate for reducing the amount of ADAMTS13 in a solution derived from plasma
An ion-exchange chromatography support for reducing the ADAMTS13 amount present in a plasma-derived solution containing human von Willebrand factor. The support includes a large-pore, vinyl polymer-type resin bearing DEAE groups, and a buffer including trisodium citrate, sodium chloride, calcium chloride, glycine and lysine.
1. A method for reducing the ADAMTS13 amount in a solution derived from human vWF-containing plasma comprising an ion-exchange chromatography step on a large-pore, vinyl polymer-type resin bearing DEAE groups, performed in a buffer comprising trisodium citrate, sodium chloride, calcium chloride, glycine and lysine.
2. The method according to claim 1 , wherein the ion-exchange chromatography support comprises a copolymer of oligoethylene glycol, glycidine methacrylate and pentaerythritol dimethacrylate on which DEAE groups are grafted.
3. The method according to claim 1 , wherein the buffer for the chromatography comprises trisodium citrate 0.01 M, calcium chloride 0.001 M, glycine 0.12 M and lysine 0.016 M and sodium chloride varying from 0.11 M to 0.17 M.
4. The method according to claim 1 , wherein the fraction which is adsorbed onto the chromatography support is eluted by increasing the sodium chloride concentration in the buffer to 0.14-0.17 M.
5. The method according to claim 1 , wherein the pH value of the buffer does range from 6.9 to 7.1.
6. The method according to claim 1 , wherein ADAMS 13 is removed by two successive chromatographic separation steps using said ion-exchange chromatography support and said buffer.
7. The method according to claim 1 , wherein the reduction in the ADAMTS13 amount is higher than 200 times as compared to the ADAMTS13 amount comprised in the initial plasma.
8. The method according to claim 1 , wherein the reduction in the ADAMTS13 amount is higher than 500 times as compared to the ADAMTS13 amount comprised in the initial plasma.