Beta‐glucans are polysaccharides of D‐glucose monomers linked by (1–3) beta‐glycosidic bonds, are found to have a potential immunogenicity risk in biotherapeutic products, and are labeled as process contaminants. A common source of beta‐glucans is from the cellulose found in traditional depth filter media. Typically, beta‐glucan impurities that leach into the product from the primary clarification depth filters can be removed by the subsequent bind‐and‐elute affinity chromatography capture step. Beta‐glucans can also be removed by a bind‐and‐elute cation exchange chromatography step, which is useful for removing beta‐glucans introduced by a post‐Protein A depth filtration step. However, the increasing prevalence of flowthrough polishing chromatography poses a challenge for beta‐glucan removal due to the lack of any bind‐and‐elute chromatography steps after the post‐Protein A depth filter. In this work, a depth filter flush strategy was developed to control beta‐glucan leaching into the product pool. Different loading conditions for the depth filtration and subsequent chromatography steps were evaluated to determine the robustness of the optimized flush strategy. Carry through runs demonstrated greater than two‐fold reduction in beta‐glucan levels using the optimized wash as compared to standard filter flush conditions.

中文翻译：

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通过改进的深层过滤冲洗策略控制从深层过滤器中浸出的 β-葡聚糖水平

β-葡聚糖是由 (1-3) β-糖苷键连接的 D-葡萄糖单体的多糖，被发现在生物治疗产品中具有潜在的免疫原性风险，并被标记为工艺污染物。β-葡聚糖的常见来源是传统深层过滤介质中的纤维素。通常，从初级澄清深度过滤器浸出到产品中的 β-葡聚糖杂质可以通过随后的结合和洗脱亲和色谱捕获步骤去除。β-葡聚糖也可以通过结合和洗脱阳离子交换色谱步骤去除，这对于去除由后蛋白质 A 深度过滤步骤引入的 β-葡聚糖很有用。然而，由于在蛋白 A 后深度过滤器之后没有任何结合和洗脱色谱步骤，流通抛光色谱法的日益普及对去除 β-葡聚糖提出了挑战。在这项工作中，开发了一种深层过滤冲洗策略来控制 β-葡聚糖浸出到产品池中。对深度过滤和后续色谱步骤的不同加载条件进行了评估，以确定优化冲洗策略的稳健性。与标准过滤器冲洗条件相比，使用优化的清洗后的连续运行表明 β-葡聚糖水平降低了两倍以上。对深度过滤和后续色谱步骤的不同加载条件进行了评估，以确定优化冲洗策略的稳健性。与标准过滤器冲洗条件相比，使用优化的清洗后的连续运行表明 β-葡聚糖水平降低了两倍以上。对深度过滤和后续色谱步骤的不同加载条件进行了评估，以确定优化冲洗策略的稳健性。与标准过滤器冲洗条件相比，使用优化的清洗后的连续运行表明 β-葡聚糖水平降低了两倍以上。