Downstream Processing Technologies/Capturing and Final Purification

Abstract

Increased pressure on upstream processes to maximize productivity has been crowned with great success, although at the cost of shifting the bottleneck to purification. As drivers were economical, focus is on now on debottlenecking downstream processes as the main drivers of high manufacturing cost. Devising a holistically efficient and economical process remains a key challenge. Traditional and emerging protein purification strategies with particular emphasis on methodologies implemented for the production of recombinant proteins of biopharmaceutical importance are reviewed. The breadth of innovation is addressed, as well as the challenges the industry faces today, with an eye to remaining impartial, fair, and balanced. In addition, the scope encompasses both chromatographic and non-chromatographic separations directed at the purification of proteins, with a strong emphasis on antibodies. Complete solutions such as integrated USP/DSP strategies (i.e., continuous processing) are discussed as well as gains in data quantity and quality arising from automation and high-throughput screening (HTS). Best practices and advantages through design of experiments (DOE) to access a complex design space such as multi-modal chromatography are reviewed with an outlook on potential future trends. A discussion of single-use technology, its impact and opportunities for further growth, and the exciting developments in modeling and simulation of DSP rounds out the overview. Lastly, emerging trends such as 3D printing and nanotechnology are covered. Graphical Abstract Workflow of high-throughput screening, design of experiments, and high-throughput analytics to understand design space and design space boundaries quickly. (Reproduced with permission from Gregory Barker, Process Development, Bristol-Myers Squibb).

Multiplex secretome engineering enhances recombinant protein production and purity

“…Thus, downstream processing has become a bottleneck in biotherapeutic production 29 . Efforts to alleviate this have mostly focused on innovations in process design to improve downstream processing capacity, speed, and economics 30,31 . It has also been suggested that cell line engineering could be used to reduce or remove problematic HCPs, lessening the need for additional purification steps 6,32 .…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…The benefits of a HT chromatography system include its availability to predict design space for dynamic binding capacity (DBC), collect data for the prediction of elution behavior, and allow significant investigation using DoE (McDonald et al, 2016). HT chromatography systems have limitations in their potential to reveal the flow distribution of process columns (Singh and Herzer, 2017). Noyes et al (2015) studied an ultra scale-down device for high-throughput depth filtration that enabled the parallel assessment of eight single-or multi-layer depth filters (∼0.2 cm 2 in cross-sectional area).…”

Critical Analysis of the Commercial Potential of Plants for the Production of Recombinant Proteins

“…Industrial processes for the extraction and purification of recombinant proteins produced by microbial and mammalian cells are well established, although downstream processing is still the main driver of overall costs (Singh and Herzer, 2018). Protein recovery is particularly straightforward when products are secreted to the culture medium by cells growing in suspension, which is generally the case for CHO cells.…”