Granted Patent B2

US 12,545,703 · App. 17/473,910 · Granted Feb 10, 2026

Inventors: Leopold Bruckschwaiger (Vienna, AT); Sonja Svatos (Berg, AT); Julia Nuernberger (Vienna, AT); Wolfgang Teschner (Vienna, AT); Harald Butterweck (Vienna, AT); Hans-Peter Schwarz (Vienna, AT); Thomas Gundinger (Vienna, AT); Bernhard Koelbl (Achau, AT); Reinhard Grausenburger (Vienna, AT); Azra Pljevljakovic (Vienna, AT)

Assignee: Takeda Pharmaceutical Company Limited

C07K1/36A61K9/0019A61K9/08A61K35/16A61K38/1709A61K39/39525A61K47/183B01D15/12B01D15/362B01D15/363B01D15/424C07K1/30C07K16/065C07K1/14C07K1/18C07K1/34

Abstract

Claims (43)

1 . A method for preparing an enriched IgG composition from plasma, the method comprising the steps of:

(a) precipitating a cryo-poor plasma fraction, in a first precipitation step, with from 6% to 10% ethyl alcohol at a pH of from 7.0 to 7.5 to form a first precipitate and a first supernatant;

(b) precipitating IgG from the first supernatant, in a second precipitation step, with from 22% to 25% ethyl alcohol at a pH of from 6.7 to 7.3 to form a second precipitate;

(c) suspending the second precipitate to form a first suspension;

(d) mixing fumed silica with the first suspension formed in (c) for at least 30 minutes; and

(e) separating a solubilized portion of the first suspension mixed with fumed silica in step (d) from a non-solubilized portion of the first suspension, thereby forming an enriched IgG composition.

2 . The method of claim 1 , wherein the second precipitation step is performed at a temperature of from −7° C. to −9° C.

3 . The method of claim 1 , wherein the second precipitate is suspended with an extraction buffer at a ratio of 1 part precipitate to from 12 parts to 18 parts of an extraction buffer.

4 . The method of claim 3 , wherein the extraction buffer has a pH of from about 4.5 to about 5.0.

5 . The method of claim 1 , wherein from 0.01 g to 0.07 g fumed silica per g of the second precipitate formed in step (b) is mixed with the first suspension in step (d).

6 . The method of claim 1 , wherein from 0.02 g to 0.06 g fumed silica per g of the second precipitate formed in step (b) is mixed with the first suspension in step (d).

7 . The method of claim 1 , wherein from 0.03 g to 0.05 g fumed silica per g of the second precipitate formed in step (b) is mixed with the first suspension in step (d).

8 . The method of claim 1 , wherein the solubilized portion of the first suspension mixed with fumed silica is separated from the non-solubilized portion of the first suspension by depth filtration.

9 . The method of claim 8 , wherein the depth filtration further comprises washing a depth filter used in the depth filtration with at least 3 filter void volumes of buffer.

10 . The method of claim 1 , wherein the soluble portion of the first suspension, separated in step (e), contains at least 85% of the IgG content of the cryo-poor plasma fraction used in step (a).

11 . The method of claim 1 , wherein the soluble portion of the first suspension, separated in step (e), contains at least 90% of the IgG content of the cryo-poor plasma fraction used in step (a).

12 . The method of claim 1 , further comprising the steps of:

(f) precipitating IgG from the solubilized portion of the suspension, in a third precipitation step, with from 22% to 28% ethyl alcohol at a pH of from 6.7 to 7.3 to form a third precipitate;

(g) suspending the third precipitate to form a second suspension;

(h) separating a solubilized portion of the second suspension formed in step (g) from a non-solubilized portion of the second suspension.

13 . The method of claim 2 , further comprising the steps of:

(f) precipitating IgG from the solubilized portion of the suspension, in a third precipitation step, with from 22% to 28% ethyl alcohol at a pH of from 6.7 to 7.3 to form a third precipitate;

(g) suspending the third precipitate to form a second suspension;

(h) separating a solubilized portion of the second suspension formed in step (g) from a non-solubilized portion of the second suspension.

14 . The method of claim 7 , further comprising the steps of:

(f) precipitating IgG from the solubilized portion of the suspension, in a third precipitation step, with from 22% to 28% ethyl alcohol at a pH of from 6.7 to 7.3 to form a third precipitate;

(g) suspending the third precipitate to form a second suspension;

(h) separating a solubilized portion of the second suspension formed in step (g) from a non-solubilized portion of the second suspension.

15 . The method of claim 12 , further comprising treating the solubilized portion of the second suspension separated in step (h) with a solvent and detergent (S/D) treatment step.

16 . The method of claim 12 , further comprising the steps of:

binding IgG in the solubilized portion of the second suspension to a cation exchange material;

eluting IgG from the cation exchange material to form a cation exchange eluate.

17 . The method of claim 16 , further comprising the steps of:

loading IgG from the cation exchange eluate onto an anion exchange column;

collecting an effluent comprising IgG from the anion exchange column to form an anion exchange flow-through.

18 . The method of claim 17 , further comprising the step of:

nanofiltering IgG from the anion exchange flow-through to form a nanofiltrate.

19 . The method of claim 18 , further comprising the step of:

ultrafiltering and diafiltering IgG from the nanofiltrate to form a filtrate having a protein concentration of at least 11% (w/v), thereby obtaining an enriched IgG composition.

20 . The method of claim 19 , wherein the ultrafiltration and diafiltration of step (n) comprises the sub-steps of:

(n1) concentrating IgG from the nanofiltrate to a protein concentration of 5±2% (w/v) to form a first IgG concentrate;

(n2) diafiltering the IgG concentrate of (n1) against a buffer comprising glycine to form an IgG diafiltrate; and

(n3) concentrating the IgG diafiltrate of (n2) to a protein concentration of at least 11% (w/v).

Assignments (4)

Priority Claims (1)

Continuity (5)

References Cited (180)