CPC C07K 1/22 (2013.01) [A61K 39/395 (2013.01); A61K 39/39591 (2013.01); C07K 1/18 (2013.01); C07K 1/34 (2013.01); C07K 1/36 (2013.01); C07K 16/00 (2013.01); C07K 16/065 (2013.01); C07K 2317/14 (2013.01); C07K 2317/31 (2013.01)] | 12 Claims |
1. A method of production of a bulk drug substance comprising a therapeutic antibody, wherein the method comprises the steps of:
(a) subjecting a harvested antibody material to Protein A chromatography;
(b) incubating the resulting Protein A eluate at a high pH;
(c) neutralizing the resulting viral inactivated solution to pH 5.5, followed by 0.2 um filtration;
(d) subjecting the neutralized viral inactivated Protein A eluate to cation exchange chromatography, followed by 0.2 um filtration;
(e) concentrating the cation exchange chromatography eluate by ultrafiltration and continuous diafiltration, followed by 0.2 um filtration;
(f) purifying the product by anion exchange chromatography in flow through mode, using membrane adsorption, followed by 0.2 um filtration;
(g) removing virus by nanofiltration;
(h) concentrating the product by ultrafiltration and continuous diafiltration into pre-formulation buffer, followed by 0.2 um filtration;
(i) adding excipients to achieve about 6 mg/mL of the product in the final formulation buffer, by mixing about 5 mM L-Histidine, about 150 mM L-Arginine Monohydrochloride, about 15% Sucrose, and about 0.06% Polysorbate 80, at pH of about 6.0, followed by 0.2 um filtration; and
(j) filling the product into sterile bags, followed by freezing and storage at −80±20° C.
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