Continuous Chromatography Device to Economically Purify Clinical-grade Antibodies

描述(申请人提供)：单抗已经成为治疗癌症的主要药物类别，超过150种抗肿瘤单抗正在生物制药管道中。大多数单抗治疗必须以4-15毫克/公斤的剂量进行，导致每个患者每年的费用高达10万美元。单抗治疗的高昂价格给患者、保险公司和医疗保健系统带来了巨大的经济负担。为了使卫生保健行业变得切实可行，必须降低这些药物的开发和生产成本。临床制造费用直接影响药物开发的总成本。蛋白质表达、细胞培养和生物反应器技术的最新发展导致了蛋白质生产和单抗效价的实质性进步。下游净化工艺的发展没有跟上步伐，是阻碍成本效益和强劲制造的瓶颈。目前，下游加工(DSP)占单抗生产总成本的70%。“行业标准”的蛋白质A捕获(PAC)是最昂贵的数字信号处理器步骤，每克产品的总成本高达40%。当前的PAC方法仍然在顺序批处理过程中使用一个大的柱。与标准批处理方法相比，连续SMB(模拟移动床)色谱具有显著的优势，包括更高效地使用昂贵的吸附剂和更小的柱、更少的缓冲液消耗以及在稳态条件下运行，从而实现更强大的过程分析。在第一阶段SBIR项目(R43 CA162632-01)中，我们研究了使用我们实验室规模的Octave(Tm)SMB系统开发连续PAC工艺纯化临床级单抗的可行性。将连续工艺与标准间歇工艺直接比较，得到了相当或更好的单抗纯度。然而，目前的Octave系统并不是为cGMP依从性或临床生产所需的规模而设计的。在第二阶段，我们建议根据Octave阀的设计和我们的第一阶段发现，开发一种大规模连续层析设备，用于经济地纯化临床级抗体。二期装置将支持流量高达2 L/分钟，在8-20小时内处理500个含有5-10 g/L单抗的L培养液，满足未来对200公斤/年治疗性单抗的需求。与批处理方法相比，第二阶段设备和优化的PAC流程将使生产率提高至少3倍，并将采用一次性流程，消除活动之间对卫生和重新验证的需要。这个第二阶段的项目将开发至少一个原型设备，并将其放置在Beta测试场。该设备将是一个可定制的即插即用的层析模块，与未来集成的连续生物处理设施兼容。该项目符合FDA 2011年的战略计划，该计划寻求开发改进的产品制造技术，包括连续工艺而不是批量方法。

DESCRIPTION (provided by applicant): Monoclonal antibodies (mAbs) have become the leading drug class for cancer treatment, and over 150 anti-neoplastic mAbs are in the biopharmaceutical pipeline. Most mAb therapies must be administered in doses of 4-15 mg/kg, resulting in per patient costs of up to $100,000 per year. The high price of mAb therapy causes a significant financial burden for patients, insurance companies and the health care system. To be- come practical for the health care industry, the cost of developing and producing these drugs must be reduced. Clinical manufacturing expenses directly contribute to the overall drug development costs. Recent developments in protein expression, cell culture, and bioreactor technologies have led to substantial advancements in protein production and mAb titers. The development of downstream purification processes has not kept pace and represents a bottleneck that impedes cost-effective and robust manufacturing. Currently, downstream processing (DSP) accounts for up to 70% of the total mAb production cost. The "industry-standard" Protein A capture (PAC) is the most expensive DSP step, contributing up to 40% of the total cost per gram of product. The current PAC method still uses one large column in a sequential batch process. Continuous SMB (simulated moving bed) chromatography offers significant advantages over standard batch methods, including more efficient use of expensive adsorbent with smaller columns, reduced buffer consumption, and operation under steady-state conditions, allowing more robust process analytics. In the Phase I SBIR project (R43 CA162632-01) we investigated the feasibility of developing a continuous PAC process for the purification of clinical-grade mAbs using our lab-scale Octave(tm) SMB System. We obtained equivalent or better mAb purity when our continuous process was directly compared with the standard batch process. How- ever, the current Octave System is not designed for cGMP compliance or the scale required for clinical manufacture. In Phase II we propose to develop a large-scale continuous chromatography device for economical purification of clinical-grade antibodies, based on the Octave valve design and our Phase I findings. The Phase II device will support flow rates up to 2 L/min and processing of 500 L culture fluid containing 5-10 g/L mAb in 8-20 hours, which matches the future demand for therapeutic mAbs in the range of 200 kg/yr. The Phase II de- vice and optimized PAC process will increase productivity at least 3-fold relative to batch methods, and will feature a single-use flow path to eliminate the need for sanitation and revalidation between campaigns. This Phase II project will develop at least one prototype device that will be placed at a Beta test site. The device will be a customizable plug-and-play chromatography module compatible with future integrated continuous bioprocessing facilities. This project fits with the 2011 FDA strategic plan, which seeks development of improved product manufacturing technologies including continuous processes rather than batch approaches.