Yamada Tsuyoshi, Yamamoto Koichi, Ishihara Takashi, Ohta Shigeru
Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd., Takasaki-shi, Gunma 370-0013, Japan; Graduate School of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin Co., Ltd., Takasaki-shi, Gunma 370-0013, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 1;1061-1062:110-116. doi: 10.1016/j.jchromb.2017.07.002. Epub 2017 Jul 3.
A depth filter is widely used in biopharmaceutical manufacturing process. In this study, we found that buffer exchange to reduce conductivity dramatically improved the removal of impurities in depth filtration. The host cell protein clearance was comparable to that of protein A affinity chromatography, which is generally used as a first capture step in monoclonal antibody purification. In addition, a combination of different depth filters showed enhanced purification. Additional flow-through purification is possible without any sample preparation, and a biopharmaceutical-quality purity level (<100ppm for host cell protein and <5% for high molecular weight species) can be attained. These results represent the possibility of an antibody manufacturing process using an entirely flow-through mode.
深层过滤器在生物制药生产过程中被广泛使用。在本研究中，我们发现通过缓冲液置换以大幅降低电导率可显著提高深层过滤中杂质的去除率。宿主细胞蛋白清除率与通常用作单克隆抗体纯化第一步捕获步骤的蛋白A亲和色谱法相当。此外，不同深层过滤器的组合显示出增强的纯化效果。无需任何样品制备即可进行额外的流通纯化，并且可达到生物制药级纯度水平（宿主细胞蛋白<100ppm，高分子量物质<5%）。这些结果表明了采用完全流通模式进行抗体生产过程的可能性。
