Claims for Patent: 10,053,489

Summary for Patent: 10,053,489

Title:	Method for purifying antibody
Abstract:	The present invention relates to a method of purifying an antibody with high purity and high quality by removing impurities by sequential use of a cation-exchange column, a culture supernatant multilayer filter and an anion-exchange column without using a protein-A column that is an affinity chromatography column which is generally used for antibody purification.
Inventor(s):	Kim; Soo Kwang (Daejeon, KR), Ahn; Yong Ho (Daejeon, KR), Kim; Young Min (Gyeonggi-do, KR), Song; Dae Hae (Daejeon, KR)
Assignee:	Prestige Biopharma PTE, LTD (SG)
Application Number:	14/896,380
Patent Claims:	1. A method for purifying an antibody, the method comprising: (a) loading a sample, which comprises the antibody and at least one host cell protein (HCP) and has a pH of 5.5-7.0 and a conductivity of 5-7 mS/cm, into an equilibrated cation-exchange column, washing the cation-exchange column, and then eluting the antibody bound to the column with elution buffer; (b) passing the eluate of step (a) through a multilayer filter, and collecting the filtrate; and (c) passing the filtrate of step (b) through an anion-exchange column, and collecting the flow-through. 2. The method according to claim 1, wherein the antibody has an isoelectric point of 7-10. 3. The method according to claim 1, wherein the antibody is Bevacizumab or Adalimumab. 4. The method according to claim 1, wherein the content of host cell protein (HCP) in the antibody eluate of step (a) is 95-99.5% lower than that in the loaded sample. 5. The method according to claim 1, wherein the cation exchange column has a functional group selected from the group consisting of carboxymethyl (CM), sulfoethyl (SE), sulfopropyl (SP), phosphate (P) and sulfonate (S). 6. The method according to claim 5, wherein the cation exchange column having sulfonate as a functional group comprises a cross-linked polymethacrylate resin. 7. The method according to claim 6, wherein step (a) comprises: (i) loading the sample comprising the antibody onto the cation exchange column equilibrated with an equilibration buffer comprising 10-50 mM phosphate (pH 5.5-7.0); (ii) washing the cation exchange column with a buffer comprising 20-40 mM sodium chloride and 10-50 mM phosphate (pH 6.0-7.0); and (iii) eluting the antibody with an elution buffer comprising 50-200 mM sodium chloride and 10-50 mM phosphate (pH 6.0-7.0). 8. The method according to claim 1, wherein the multilayer filter has a product water conductivity of 8.66-10.5 .mu.S/cm or 19.34-53.2 .mu.S/cm post autoclave and pure water flush of 10 liters per ft.sup.2 (100 L/m.sup.2) of surface area of the multilayer filter. 9. The method according to claim 1, wherein the antibody eluate of step (b) is subjected to viral inactivation before it is passed through the multilayer filter. 10. The method according to claim 1, wherein the antibody eluate of step (b) is adjusted to a pH of 5.5-7.0 before it is passed through the multilayer filter. 11. The method according to claim 1, wherein the content of host cell protein (HCP) in the filtrate of step (b) is 1,200-2,500 times lower than that in the antibody eluate of step (a). 12. The method according to claim 1, wherein the anion exchange column in step (c) has a functional group of a quartenary amine and further comprises a cross-linked agarose base matrix. 13. The method according to claim 1, wherein the content of host cell protein (HCP) in the protein purified by the method is 0.001-10 ppm. 14. The method according to claim 1, wherein the method comprises no additional column chromatography step.

Details for Patent 10,053,489

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	12/31/2002		2033-06-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	02/21/2008		2033-06-05
Abbvie Inc.	HUMIRA	adalimumab	Injection	125057	04/24/2013		2033-06-05