Victor Pasquier, Kevin Botelho Ferreira, Morgane Lergenmuller, Alexis Tottoli, Arnaud Perilleux, Jonathan Souquet, Jean-Marc Bielser
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引用次数: 0
摘要
膜层析装置是填料床树脂的可行替代品,可实现单克隆抗体和 Fc 融合蛋白的高产纯化级联。本研究评估了离子交换和蛋白 A 膜层析的性能,并将其与同类树脂进行了比较。其中两种测试膜的蛋白 A 动态结合能力高于 50 克/升,停留时间为 0.2 分钟。在抛光方面,与具有类似配体的树脂相比,聚合体清除率通常低于膜分离性能。然而,膜的产量和生产率的提高足以让我们考虑使用膜。此外,寿命研究表明,膜的性能在循环过程中保持稳定。大多数测试膜都达到了一百次循环,对工艺性能和产品质量没有影响。最后,从捕获到抛光,纯化级联完全用膜操作,宿主细胞蛋白质(小于 50 ppm)和聚集物(等于或小于 1%)达到良好水平。这项研究结果表明,在单克隆抗体和 Fc 融合蛋白纯化过程中,膜完全可以取代树脂色谱法。
Assessment of membrane-based downstream purification processes as a replacement to traditional resin bead for monoclonal antibody purification.
Membrane chromatography devices are a viable alternative to packed-bed resins and enable highly productive purification cascades for monoclonal antibodies and Fc-fusion proteins. In this study, ion exchange and protein A membrane chromatography performances were assessed and compared with their resin counterparts. Protein A dynamic binding capacities were higher than 50 g/L for two of the tested membranes and with a residence time of 0.2 min. For polishing, it was observed that aggregate clearance was generally less performant with membrane separation when compared to resins with similar ligands. However, the comparable yield and increased productivity of membranes could be enough to consider their implementation. In addition, lifetime studies demonstrated that the performance of membranes remained robust over cycles. One hundred cycles were reached for most of the tested membranes with no impact on the process performance nor product quality. Finally, purification cascades were fully operated with membranes, from capture to polishing, reaching good levels of host cells proteins (less than 50 ppm) and aggregates (equal to or less than 1%). The outcome of this study demonstrated that resin chromatography could be fully replaced by membranes for monoclonal antibody and Fc-fusion protein purification processes.
期刊介绍:
Biotechnology Progress , an official, bimonthly publication of the American Institute of Chemical Engineers and its technological community, the Society for Biological Engineering, features peer-reviewed research articles, reviews, and descriptions of emerging techniques for the development and design of new processes, products, and devices for the biotechnology, biopharmaceutical and bioprocess industries.
Widespread interest includes application of biological and engineering principles in fields such as applied cellular physiology and metabolic engineering, biocatalysis and bioreactor design, bioseparations and downstream processing, cell culture and tissue engineering, biosensors and process control, bioinformatics and systems biology, biomaterials and artificial organs, stem cell biology and genetics, and plant biology and food science. Manuscripts concerning the design of related processes, products, or devices are also encouraged. Four types of manuscripts are printed in the Journal: Research Papers, Topical or Review Papers, Letters to the Editor, and R & D Notes.