Highly efficient anti-TFPI antibody composition
The present disclosure relates to a pharmaceutical highly efficient anti-TFPI antibody composition including an anti-TFPI antibody for treating hemophilia, in which the content of HCP in a drug substance is less than 10.0 ng/mg, and the content of LPA in the drug substance or the drug product is less than 1.0 ng/mg. According to the present disclosure, a highly efficient anti-TFPI antibody composition having very low contents of HCP and LPA may be provided by providing a separation/purification process capable of minimizing anti-TFPI antibody polymer generation, and may be effectively used for the treatment of antibody-induced hemophilia patients and for the prevention of blood coagulation disease.
1. A method of preparing a pharmaceutical composition comprising a purified anti-tissue factor pathway inhibitor (anti-TFPI) antibody, wherein a content of host cell protein (HCP) in the composition is less than 10.0 ng/mg, and a content of leachable protein A (LPA) in the composition is less than 1.0 ng/mg,
wherein the anti-TFPI antibody is an IgG4 antibody,
the method comprising
(a) a step of clarifying a culture containing the anti-TFPI antibody to obtain a clarified culture, wherein the clarified culture is subjected to following (b)-(g), in this order;
(b) an affinity chromatography process at pH of 3.0 to 3.8, which further comprises the steps of:
(b-1) neutralizing an eluate, which passed through an affinity chromatography column, in the range of pH 4.0 to 6.0; and
(b-2) leaving the neutralized eluate obtained in (b-1) to stand at a temperature of 15 to 25° C. for 8 to 16 hours;
(c) a viral inactivation process;
(d) an ultrafiltration and diafiltration process;
(e) a mixed-mode chromatography process;
(f) a nano-filtration process; and
(g) a concentration process,
wherein the mixed-mode chromatography process (e) comprises performing two or more plural mixed-mode chromatography processes using different types of column resin to remove the anti-TFPI antibody in a form of polymer.
2. The method of claim 1 , wherein a total content of the HCP and the LPA in the composition is 5 ng/mg or less.
3. The method of claim 1 , wherein the mixed-mode chromatography process (e) removes 98% or more of the anti-TFPI antibody in a form of polymer.
4. The method of claim 1 , wherein the concentration process comprises a rinse solution circulation/recovery process.
5. The method of claim 1 , wherein a content of the anti-TFPI antibody in a form of polymer in the composition is less than 5%.
6. The method of claim 1 , wherein a content of the anti-TFPI antibody in the composition, which is in a form of monomer, is greater than 60% based on a total amount of the anti-TFPI antibody contained in the composition.
7. The method of claim 1 , wherein the HCP content in a composition obtained after a purification comprising (b)-(g) is at least 50,000-fold lower than the HCP content in the clarified culture obtained in (a) and before the purification.
8. The method of claim 1 , wherein a relative potency value of the composition is 85% or higher compared to that of the composition in which 100% of the anti-TFPI antibody is in a form of monomer.