Department of Chemical and Biochemical Engineering, Missouri University of Science and Technology, Rolla, Missouri.
Biotechnol Bioeng. 2020 Aug;117(8):2588-2609. doi: 10.1002/bit.27362. Epub 2020 May 16.
Endotoxins are the major contributors to the pyrogenic response caused by contaminated pharmaceutical products, formulation ingredients, and medical devices. Recombinant biopharmaceutical products are manufactured using living organisms, including Gram-negative bacteria. Upon the death of a Gram-negative bacterium, endotoxins (also known as lipopolysaccharides) in the outer cell membrane are released into the lysate where they can interact with and form bonds with biomolecules, including target therapeutic compounds. Endotoxin contamination of biologic products may also occur through water, raw materials such as excipients, media, additives, sera, equipment, containers closure systems, and expression systems used in manufacturing. The manufacturing process is, therefore, in critical need of methods to reduce and remove endotoxins by monitoring raw materials and in-process intermediates at critical steps, in addition to final drug product release testing. This review paper highlights a discussion on three major topics about endotoxin detection techniques, upstream processes for the production of therapeutic molecules, and downstream processes to eliminate endotoxins during product purification. Finally, we have evaluated the effectiveness of endotoxin removal processes from a perspective of high purity and low cost.
内毒素是由污染的药物产品、制剂成分和医疗器械引起的发热反应的主要贡献者。重组生物制药产品是使用包括革兰氏阴性菌在内的活体生物制造的。革兰氏阴性菌死亡后，外细胞膜中的内毒素（也称为脂多糖）会释放到裂解物中，在那里它们可以与生物分子相互作用并形成键，包括靶治疗化合物。生物制品的内毒素污染也可能通过水、赋形剂等原材料、培养基、添加剂、血清、设备、容器封闭系统和用于制造的表达系统发生。因此，除了最终的药物产品放行测试外，制造过程迫切需要通过监测原材料和关键步骤中的中间过程来减少和去除内毒素的方法。本文综述了关于内毒素检测技术、治疗分子生产的上游工艺以及产品纯化过程中去除内毒素的下游工艺等三个主要主题的讨论。最后，我们从高纯度和低成本的角度评估了内毒素去除工艺的有效性。
