Method for filtering fibrinogen
Disclosed is a method for filtering a fibrinogen composition, including the following steps: a) purifying the fibrinogen composition by chromatographic purification using an elution buffer comprising arginine; b) optionally, at least one step of filtering the fibrinogen composition obtained by chromatographic elution in step a), on a filter having a pore size of between 0.08 μm and 0.22 μm, c) filtering the fibrinogen composition obtained by chromatographic elution in step a), or optionally obtained in step b), on a symmetrical filter having a pore size of between 15 nm and 25 nm, and preferably between 18 nm and 22 nm, and d) recovering the resulting fibrinogen solution, the filtering method being carried out without adding arginine after step a), at a high capacity and without a prior freezing and/or thawing step.
1 . A process for filtering a fibrinogen composition, comprising the following steps:
a) chromatographic purification of the fibrinogen composition with an elution buffer comprising arginine;
b) performing at least one filtraton step of the fibrinogen composition obtained by elution chromatography in step a) on a symmetrical filter having a pore size between 15 nm and 25 nm, and
c) recovering the resulting fibrinogen solution,
wherein the elution buffer used in step a) comprises an arginine concentration of at least 200 mM, and
wherein the fibrinogen yield of the filtration is higher than 90% said filtration process being carried out without addition of arginine after step a).
2 . The process of claim 1 , further comprising at least one additional filtration step performed prior to the symmetrical filtration step, comprising filtering the fibrinogen on a filter having a pore size between 0.08μm and 0.22 μm.
3 . The process of claim 1 , further comprising at least one additional filtration step performed prior to the symmetrical filtration step, comprising filtering the fibrinogen comprising on a filter having a pore size between 0.08 um and 0.22 um; filtering the fibrinogen solution obtained in step b) on a filter having a pore diameter between 15 nm and 50 nm.
4 . The process according to claim 2 , wherein the symmetrical filter has a pore size between 18 nm and 22 nm.
5 . The process according to claim 1 , wherein the fibrinogen composition is obtained from plasma fractions, milk of transgenic animals, or cell culture supernatant.
6 . The process according to claim 1 , wherein the chromatographic purification step is performed by affinity, mixed-mode, or ion-exchange chromatography.
7 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration between 200 mM and 800 mM.
8 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration between 200 mM and 600 mM.
9 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration between 200 mM and 400 mM.
10 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration of at least 200 mM and said filtration process is carried out at a capacity of at least 0.25 kg/m 2 .
11 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration of at least 200 mM and said filtration process is carried out at a capacity of at least 0.35 kg/m 2 .
12 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration of at least 400 mM and said filtration process is carried out at a capacity of at least 0.25 kg/m 2 .
13 . The process according to claim 1 , wherein the elution buffer used in step a) comprises an arginine concentration of at least 400 mM and said filtration process is carried out at a capacity of at least 0.35 kg/m 2 .
14 . The process of claim 1 , further comprising at least one additional filtration step performed prior to the symmetrical filtration step, comprising filtering the fibrinogen on a filter having a pore size between 0.15 μm and 0.22 μm, and then filtering on a filter having a pore size between 0.08 μm and 0.15 μm.
15 . The process according to claim 14 , wherein the first filtration is carried out on a filter having a pore size of 0.2 μm, and the second filtration is carried out on a filter having a pore size of 0.1 μm.
16 . The process according to claim 1 , wherein said filtration process is being carried out without addition of arginine after step a), at a capacity of at least 0.2 kg fibrinogen/m 2 , and said fibrinogen composition not being previously frozen and/or thawed.