Integration of a perfusion reactor and continuous precipitation in an entirely membrane-based process for antibody capture

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Engineering in Life Sciences Pub Date : 2023-09-07 DOI:10.1002/elsc.202300219
Gabriele Recanati, Magdalena Pappenreiter, Christoph Gstoettner, Patrick Scheidl, Elena Domínguez Vega, Bernhard Sissolak, Alois Jungbauer
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Abstract

Continuous precipitation coupled with continuous tangential flow filtration is a cost-effective alternative for the capture of recombinant antibodies from crude cell culture supernatant. The removal of surge tanks between unit operations, by the adoption of tubular reactors, maintains a continuous harvest and mass flow of product with the advantage of a narrow residence time distribution (RTD). We developed a continuous process implementing two orthogonal precipitation methods, CaCl2 precipitation for removal of host-cell DNA and polyethylene glycol (PEG) for capturing the recombinant antibody, with no influence on the glycosylation profile. Our lab-scale prototype consisting of two tubular reactors and two stages of tangential flow microfiltration was continuously operated for up to 8 days in a truly continuous fashion and without any product flow interruption, both as a stand-alone capture and as an integrated perfusion-capture. Furthermore, we explored the use of a negatively charged membrane adsorber for flow-through anion exchange as first polishing step. We obtained a product recovery of approximately 80% and constant product quality, with more than two logarithmic reduction values (LRVs) for both host-cell proteins and host-cell DNA by the combination of the precipitation-based capture and the first polishing step.

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在完全基于膜的抗体捕获过程中整合灌注反应器和连续沉淀。
连续沉淀与连续切向流过滤相结合是从粗细胞培养上清液中捕获重组抗体的一种具有成本效益的替代方案。通过采用管式反应器,在单元操作之间移除缓冲罐,保持了产品的连续收获和质量流,具有窄停留时间分布(RTD)的优点。我们开发了一种实施两种正交沉淀方法的连续工艺,CaCl2沉淀用于去除宿主细胞DNA,聚乙二醇(PEG)用于捕获重组抗体,对糖基化谱没有影响。我们的实验室规模的原型由两个管式反应器和两级切向流微滤组成,以真正连续的方式连续运行长达8天,没有任何产品流中断,无论是作为独立捕获还是作为集成灌注捕获。此外,我们探索了使用带负电荷的膜吸附器进行流通式阴离子交换作为第一抛光步骤。通过基于沉淀的捕获和第一抛光步骤的组合,我们获得了约80%的产品回收率和恒定的产品质量,宿主细胞蛋白质和宿主细胞DNA都具有两个以上的对数还原值(LRV)。
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来源期刊
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
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