Ishii Yoichi, Tsukahara Masayoshi, Wakamatsu Kaori
Bio-process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki-shi, Gunma 370-0013, Japan; Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu-shi, Gunma 376-8515, Japan.
Kyowa Hakko Kirin Co., Ltd., 1-6-1 Ohte-machi, Chiyoda-ku, Tokyo 100-8185, Japan.
J Biosci Bioeng. 2016 Apr;121(4):464-70. doi: 10.1016/j.jbiosc.2015.08.002. Epub 2015 Oct 20.
The goal of developing a monoclonal antibody (mAb) production process is high productivity and high quality. Because the productivity and quality of mAbs depend on cell line properties, the selection of cell lines suitable for large-scale production is an important stage in process development for mAb production. The light chain (LC) is important for antibody folding and assembly in the endoplasmic reticulum; cell lines that secrete a large amount of LCs in the medium secrete high-quality antibodies with high productivity. LC contents in culture media have been estimated by western blotting, reverse-phase high-performance liquid chromatography, and enzyme-linked immunosorbent assay. However, these analyses require fine tuning of experimental conditions for each antibody analyzed. Here we report a rapid and simple high-sensitivity size-exclusion chromatography (HS-SEC) method to evaluate the contents of low-molecular weight species (LMWS, mainly consisting of LC monomers and dimers) and high-molecular weight species (HMWS, aggregates) in the media for cell line screening. Because LMWS and HMWS are important indicators of productivity and quality, respectively, for cell line screening, HS-SEC will be useful in the first step of cell line selection needed for large-scale production.
开发单克隆抗体（mAb）生产工艺的目标是实现高生产率和高质量。由于单克隆抗体的生产率和质量取决于细胞系的特性，因此选择适合大规模生产的细胞系是单克隆抗体制备工艺开发中的一个重要阶段。轻链（LC）对于内质网中的抗体折叠和组装很重要；在培养基中分泌大量轻链的细胞系能够分泌出具有高生产率的高质量抗体。培养基中的轻链含量已通过蛋白质印迹法、反相高效液相色谱法和酶联免疫吸附测定法进行估算。然而，这些分析需要针对每种分析的抗体对实验条件进行微调。在此，我们报告一种快速简便的高灵敏度尺寸排阻色谱法（HS-SEC），用于评估培养基中低分子量物质（LMWS，主要由轻链单体和二聚体组成）和高分子量物质（HMWS，聚集体）的含量，以用于细胞系筛选。由于低分子量物质和高分子量物质分别是细胞系筛选中生产率和质量的重要指标，高灵敏度尺寸排阻色谱法在大规模生产所需的细胞系选择的第一步中将很有用。
