Manufacture of factor H (FH) and FH-derivatives from plasma
The present invention provides compositions and pharmaceutical formulations of Factor H derived from plasma. Also provided are methods for the manufacture of the Factor H compositions and formulations, as well as methods for the treatment of diseases associated with Factor H dysfunction.
1. A method for preparing an enriched Factor H composition from plasma, the method comprising the steps of:
(a) extracting Factor H from a Fraction I precipitate and/or a Fraction II+III precipitate with an extraction buffer selected from the group consisting of: a solution comprising from about 10 mM to about 250 mM of a buffering agent, a conductivity of from about 5 mS/cm to about 100 mS/cm, and a pH from about 6.0 to about 9.0, and a solution comprising from about 2.5 mM to about 100 mM of a buffering agent, a conductivity of from about 0.5 mS/cm to about 2.0 mS/cm, and a pH from about 4.0 to about 5.5, to form a Factor H extract; and
(b) precipitating an impurity from the Factor H extract to form a supernatant containing Factor H, thereby preparing an enriched Factor H composition.
2. The method of claim 1 , wherein precipitation step (b) comprises precipitating an impurity from the Factor H extract using a final alcohol concentration of from 10% to 20% at a pH of from 7.0 to 9.0.
3. The method of claim 1 , wherein precipitation step (b) comprises precipitating an impurity from the Factor H extract using a final alcohol concentration of from 14% to 16% at a pH of from 7.5 to 8.5.
4. The method of claim 1 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0,
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0.
5. The method of claim 1 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 24% to 26% at a pH of from 5.8 to 6.2.
6. The method of claim 2 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0.
7. The method of claim 2 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 24% to 26% at a pH of from 5.8 to 6.2.
8. The method of claim 3 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 20% to 30% at a pH of from 5.0 to 7.0.
9. The method of claim 3 , wherein the method further comprises the step of:
(c) precipitating Factor H from the supernatant formed in step (b) using a final alcohol concentration of from 24% to 26% at a pH of from 5.8 to 6.2.
10. The method of claim 1 , wherein the method further comprises the steps of:
(c) binding Factor H to an anion exchange resin; and
(d) eluting Factor H from the anion exchange resin with an elution buffer, thereby forming a first eluate containing Factor H.
11. The method of claim 10 , further comprising the steps of:
(e) binding Factor H from the first eluate to a heparin affinity resin; and
(f) eluting the Factor H from the heparin affinity resin with an elution buffer, thereby forming a second eluate containing Factor H.
12. The method of claim 1 , further comprising the steps of:
(i) binding an impurity to an anion exchange resin under conditions such that Factor H does bind to the resin and is collected in the flow through fraction;
(ii) binding Factor H from the flow through fraction to a heparin affinity resin;
(iii) eluting Factor H from the heparin affinity resin to form an eluate;
(iv) binding Factor H from the eluate to an anion exchange resin; and
(v) eluting Factor H from the anion exchange resin, thereby further purifying Factor H.
13. The method of claim 6 , wherein the method further comprises the steps of:
(d) binding Factor H to an anion exchange resin; and
(e) eluting Factor H from the anion exchange resin with an elution buffer, thereby forming a first eluate containing Factor H.
14. The method of claim 13 , further comprising the steps of:
(f) binding Factor H from the first eluate to a heparin affinity resin; and
(g) eluting the Factor H from the heparin affinity resin with an elution buffer, thereby forming a second eluate containing Factor H.
15. The method of claim 6 , further comprising the steps of:
(i) binding an impurity to an anion exchange resin under conditions such that Factor H does bind to the resin and is collected in the flow through fraction;
(ii) binding Factor H from the flow through fraction to a heparin affinity resin;
(iii) eluting Factor H from the heparin affinity resin to form an eluate;
(iv) binding Factor H from the eluate to an anion exchange resin; and
(v) eluting Factor H from the anion exchange resin, thereby further purifying Factor H.
16. The method of claim 1 , further comprising at least one viral inactivation step.
17. A method for treating a disease associated with Factor H dysfunction in a subject in need thereof, the method comprising administering a therapeutically effective dose of a pharmaceutical composition of Factor H prepared according to the method of claim 1 .