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Achieving High Mass-Throughput of Therapeutic Proteins Through Parvovirus Retentive Filters

机译：通过细小病毒保持过滤器实现治疗蛋白的高通量

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Parvovirus retentive filters that assure removal of viruses and virus-like particles during the production of therapeutic proteins significantly contribute to total manufacturing costs. Operational approaches that can increase throughput and reduce filtration area would result in a significant cost savings. A combination of methods was used to achieve high throughputs of an antibody or therapeutic protein solution through three parvovirus retentive filters. These methods included evaluation of diatomaceous earth or size-based prefilters, the addition of additives, and the optimization of protein concentration, temperature, buffer composition, and solution pH. An optimum temperature of 35°C was found for maximizing throughput through the Virosart CPV and Viresolve Pro filters. Mass-throughput values of 7.3, 26.4, and 76.2 kg/m~2 were achieved through the Asahi Planova 20N, Virosart CPV, and Viresolve Pro filters, respectively, in 4 h of processing. Mass-throughput values of 73, 137, and 192 kg/m2 were achieved through a Millipore Viresolve Pro filter in 4.0, 8.8, and 22.1 h of processing, respectively, during a single experiment. However, large-scale parvovirus filtration operations are typically controlled to limit volumetric throughput to below the level achieved during small-scale virus spiking experiments. The virus spike may cause significant filter plugging, limiting throughput. Therefore newer parvovirus filter spiking strategies should be adopted that may lead to more representative viral clearance data and higher utilization of large-scale filter capacity.
机译：细小病毒保留过滤器可确保在生产治疗性蛋白质的过程中去除病毒和类病毒颗粒，从而大大降低了总制造成本。可以提高生产量并减少过滤面积的操作方法将节省大量成本。通过三种细小病毒保持过滤器，使用多种方法组合可实现高通量的抗体或治疗性蛋白溶液。这些方法包括评估硅藻土或基于尺寸的预滤器，添加添加剂以及优化蛋白质浓度，温度，缓冲液组成和溶液pH。找到了35°C的最佳温度，以通过Virosart CPV和Viresolve Pro过滤器最大化产量。在处理4小时后，通过Asahi Planova 20N，Virosart CPV和Viresolve Pro过滤器分别获得了7.3、26.4和76.2 kg / m〜2的质量通量值。在一个实验中，通过Millipore Viresolve Pro过滤器分别在4.0、8.8和22.1 h的处理中实现了73、137和192 kg / m2的质量通量值。但是，通常控制细小病毒的大规模过滤操作，以将体积通量限制在小规模病毒加标实验中所达到的水平以下。病毒峰值可能会导致严重的过滤器堵塞，从而限制吞吐量。因此，应采用更新的细小病毒滤池加标策略，这可能会导致更具代表性的病毒清除率数据和更高的大规模滤池容量利用率。

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