β-Glucan process-related impurities can be introduced into biopharmaceutical products via upstream or downstream processing or via excipients. This study obtained a comprehensive process-mapping dataset for five monoclonal antibodies to assess β-glucan introduction and clearance during development and production runs at various scales. Overall, 198 data points were available for analysis. The greatest β-glucan concentrations were found in the depth-filtration filtrate (37–2,745 pg/ml). Load volume correlated with β-glucan concentration in the filtrate, whereas flush volume was of secondary importance. Cation-exchange chromatography significantly cleared β-glucans. Furthermore, β-glucan leaching from the Planova 20N virus removal filter was reduced by increasing the flush volume (1 vs. 10 L/m²). β-glucan concentrations after filter flush with 10 L/m² were consistently <10 pg/ml. No or only limited β-glucan clearance was attained via ultrafiltration/diafiltration (UF/DF). However, during the first run with monoclonal antibody (mAb) 4, β-glucan concentration in the UF/DF retentate was 10.8 pg/mg, potentially due to β-glucan leaching from the first run with a regenerated cellulose membrane. Overall, β-glucan levels in the final mAb drug substance were 1–12 pg/mg. Assuming high doses of 1,000–5,000 mg, a β-glucan contamination at 20 pg/mg would translate to 20–100 ng/dose, which is below the previously suggested threshold for product safety (≤500 ng/dose).

中文翻译：

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单克隆抗体下游加工中β-葡聚糖的引入和清除

β-葡聚糖工艺相关杂质可以通过上游或下游加工或通过赋形剂引入生物制药产品。本研究获得了五种单克隆抗体的综合过程映射数据集，以评估在各种规模的开发和生产过程中 β-葡聚糖的引入和清除。总体而言，有 198 个数据点可用于分析。在深度过滤滤液中发现了最高的 β-葡聚糖浓度 (37–2,745 pg/ml)。上样量与滤液中的 β-葡聚糖浓度相关，而冲洗量是次要的。阳离子交换色谱法显着清除了β-葡聚糖。此外，通过增加冲洗量（1 对 10 L/m ²）。^{用 10 L/m 2}冲洗过滤器后的 β-葡聚糖浓度始终小于 10 pg/ml。通过超滤/渗滤 (UF/DF) 没有或仅获得有限的 β-葡聚糖清除。然而，在第一次使用单克隆抗体 (mAb) 4 运行期间，UF/DF 截留物中的 β-葡聚糖浓度为 10.8 pg/mg，这可能是由于第一次使用再生纤维素膜时 β-葡聚糖的浸出。总体而言，最终 mAb 药物物质中的 β-葡聚糖水平为 1-12 pg/mg。假设 1,000–5,000 mg 的高剂量，20 pg/mg 的 β-葡聚糖污染将转化为 20–100 ng/剂量，低于先前建议的产品安全阈值（≤500 ng/剂量）。