Immunoglobulin reduced in thrombogenic agents and preparation thereof
The invention relates to an immunoglobulin composition reduced in thrombogenic agents and to methods for its preparation. One method comprises subjecting an immunoglobulin containing solution to a negative cation exchanger chromatography at a pH in the range of higher than 3.8 to equal to or lower than 5.3. The solution can also be subjected to a negative anion exchanger chromatography at a pH in the range of 7 to 8.2.
1. A method for preparing an immunoglobulin composition comprising the steps of:
a) contacting an immunoglobulin solution obtained by Cohn Fractionation and/or Kistler-Nitschmann fractionation or a fraction thereof with an anion exchanger equilibrated at a pH in the range of 7 to 8.2, and a cation exchanger equilibrated at a pH in the range of higher than 3.8 to equal to or lower than 5.3 to allow prekallikrein activator (PKA) (“PKA”) to bind to the anion exchanger, and Factor XIa, Factor XI and/or Kallikrein to bind to the cation exchanger; and
b) collecting the unbound fractions comprising immunoglobulin.
2. The method according to claim 1 , wherein the cation exchanger chromatography is carried out twice.
3. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of higher than 3.8 to equal to or lower than 5.0.
4. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of equal to or higher than 4.0 to equal to or lower than 5.0.
5. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of higher than 3.8 to equal to or lower than 4.7.
6. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH of about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, or 4.7.
7. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of higher than 3.8 to equal to or lower than 4.3.
8. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of equal to or higher than 4.0 to equal to or lower than 4.3.
9. The method according to claim 1 , wherein the cation exchanger chromatography is carried out at a pH in the range of equal to or higher than 4.1 to equal to or lower than 4.3.
10. The method according to claim 1 , further comprising a negative chromatography using a chromatographic material comprising three-dimensional cross-linked hydrophobic acrylic polymer.
11. The method according to claim 1 , wherein the cation exchanger is in the form of a membrane.
12. The method according to claim 1 , wherein the cation exchanger comprises a sulfonic functional group.
13. An immunoglobulin composition derived from blood or blood fractions, comprising a total protein level of 4%-10% and that is obtained according to the method of claim 1 .
14. A receptacle containing the immunoglobulin composition according to claim 13 .