Process for reducing subvisible particles in a pharmaceutical formulation
The present disclosure provides a stable protein composition containing a surfactant and having less than 400 subvisible particles of 10 microns or greater diameter per container, or less than 10,000 subvisible particles of 2 microns or greater per container. A method of manufacturing such a stable protein composition is disclosed, which includes a unit of operation that removes or decreases an esterase activity that degrades the surfactant. The unit of operation may be hydrophobic interaction chromatography or filtration, mixed mode chromatography, or the like.
1. A process for manufacturing a formulated drug substance comprising an antibody produced by a host cell, the process comprising:
in a first purification step, separating the antibody and a phospholipase B-like 2 protein (PLBL2) from the host cell to obtain a composition comprising the antibody and the PLBL2, wherein the first purification step includes protein affinity capture;
in a second purification step, separating the antibody from the PLBL2 by contacting the composition with one of a hydrophobic interaction chromatography (HIC) resin or an HIC membrane; and
after the second purification step, combining the antibody and a fatty acid ester to make the formulated drug substance, wherein the formulated drug substance is essentially free of subvisible particles.
2. The process of claim 1 , wherein the first purification step further includes an ion exchange process.
3. The process of claim 1 , wherein the one of the HIC resin or the HIC membrane preferentially binds the PLBL2 and does not bind the antibody.
4. The process of claim 1 , wherein the one of the HIC resin or the HIC membrane comprises a hydrophobic moiety selected from the group consisting of a phenyl, ether, butyl, octyl, straight chain alkane, branch chain alkane, 8-carbon alkane, and 18-carbon alkane.
5. The process of claim 1 , further comprising:
concentrating the antibody after separating the antibody from the PLBL2.
6. The process of claim 5 , further comprising:
combining a buffer and a thermal stabilizer with the concentrated antibody.
7. The process of claim 1 , further comprising:
storing the formulated drug substance at 5° C. for at least 6 months.
8. The process of claim 1 , further comprising:
disposing a volume of the formulated drug substance in a container, wherein the volume of the formulated drug substance contains less than 400 particles having a diameter of ≥10 microns.
9. The process of claim 8 , wherein the container contains less than 10,000 particles having a diameter of ≥2 microns.
10. The process of claim 1 , wherein the antibody is a recombinant human monoclonal antibody.
11. The process of claim 1 , wherein the PLBL2 comprises the amino acid sequence of SEQ ID NO:1.
12. The process of claim 1 , wherein the fatty acid ester is a polyoxyethylene (20) sorbitan ester.
13. The process of claim 12 , wherein the polyoxyethylene (20) sorbitan ester is polysorbate 20 or polysorbate 80.
14. The process of claim 1 , wherein the concentration of the antibody in the formulated drug substance is at least 30 mg/mL.
15. The process of claim 1 , further comprising storing the formulated drug substance at 5° C. for at least 6 months, after which the formulated drug substance includes less than 400 particles having an average mean diameter of ten microns or more.
16. The process of claim 1 , further comprising storing the formulated drug substance at 5° C. for at least 6 months, after which the formulated drug substance includes less than 10,000 particles having an average mean diameter of 2 microns or more.
17. A process for manufacturing a formulated drug substance comprising an IgG antibody produced by a host cell, the process comprising:
in a first purification step, separating the IgG antibody and a phospholipase B-like 2 protein (PLBL2) from the host cell to obtain a composition comprising the IgG antibody and the PLBL2, wherein the first purification step comprises performing protein affinity capture and an ion exchange process;
in a second purification step, contacting the composition with one of a hydrophobic interaction chromatography (HIC) resin or an HIC membrane to preferentially bind the PLBL2 without binding the IgG antibody;
after the second purification step, combining the IgG antibody, a fatty acid ester, a buffer, and a thermal stabilizer to make the formulated drug substance; and
storing the formulated drug substance at 5° C. for at least 6 months, after which the formulated drug substance includes less than 400 particles having an average mean diameter of ten microns or more.
18. The process of claim 17 , wherein after storage the formulated drug substance includes less than 10,000 particles having an average mean diameter of 2 microns or more.
19. The process of claim 17 , wherein the ion exchange process comprises performing anion exchange chromatography.
20. The process of claim 17 , wherein the ion exchange process comprises performing cation exchange chromatography.
21. The process of claim 17 , wherein the host cell is a Chinese hamster ovary cell.