Perfusion Cell Culture Decreases Process and Product Heterogeneity in a Head-to-Head Comparison With Fed-Batch.

In this study, the authors compared the impacts of fed-batch and perfusion platforms on process and product attributes for IgG1- and IgG4-producing cell lines. A "plug-and-play" approach is applied to both platforms at bench scale, using commercially available basal and feed media, a standard feed strategy for fed-batch and ATF filtration for perfusion. Product concentration in fed-batch is 2.5 times greater than perfusion, while average productivity in perfusion is 7.5 times greater than fed-batch. PCA reveals more variability in the cell environment and metabolism during the fed-batch run. LDH measurements show that exposure of product to cell lysate is 7-10 times greater in fed-batch. Product analysis shows larger abundances of neutral species in perfusion, likely due to decreased bioreactor residence times and extracellular exposure. The IgG1 perfusion product also has higher purity and lower half-antibody. Glycosylation is similar across both culture modes. The first perfusion harvest slice for both product types shows different glycosylation than subsequent harvests, suggesting that product quality lags behind metabolism. In conclusion, process and product data indicate that intra-lot heterogeneity is decreased in perfusion cultures. Additional data and discussion is required to understand the developmental, clinical and commercial implications, and in what situations increased uniformity would be beneficial.

Reminder Cards Improve Physician Documentation of Obesity But Not Obesity Counseling.

BACKGROUND AND OBJECTIVES: Physicians frequently fail to document obesity and obesity-related counseling. We sought to determine whether attaching a physical reminder card to patient encounter forms would increase electronic medical record (EMR) assessment of and documentation of obesity and dietary counseling. METHODS: Reminder cards for obesity documentation were attached to encounter forms for patient encounters over a 2-week intervention period. For visits in the intervention period, the EMR was retrospectively reviewed for BMI, assessment of "obesity" or "morbid obesity" as an active problem, free-text dietary counseling within physician notes, and assessment of "dietary counseling" as an active problem. These data were compared to those collected through a retrospective chart review during a 2-week pre-intervention period. We also compared physician self-report of documentation via reminder cards with EMR documentation. RESULTS: We found significant improvement in the primary endpoint of assessment of "obesity" or "morbid obesity" as an active problem (42.5% versus 28%) compared to the pre-intervention period. There was no significant difference in the primary endpoints of free-text dietary counseling or assessment of "dietary counseling" as an active problem between the groups. Physician self-reporting of assessment of "obesity" or "morbid obesity" as an active problem (77.7% versus 42.5%), free-text dietary counseling on obesity (69.1% versus 35.4%) and assessment of "dietary counseling" as an active problem (54.3% versus 25.2%) were all significantly higher than those reflected in EMR documentation. CONCLUSIONS: This study demonstrates that physical reminder cards are a successful means of increasing obesity documentation rates among providers but do not necessarily increase rates of obesity-related counseling or documentation of counseling. Our study suggests that even with such interventions, physicians are likely under-documenting obesity and counseling compared to self-reported rates.

The business impact of an integrated continuous biomanufacturing platform for recombinant protein production.

The biotechnology industry primarily uses batch technologies to manufacture recombinant proteins. The natural evolution of other industries has shown that transitioning from batch to continuous processing can yield significant benefits. A quantitative understanding of these benefits is critical to guide the implementation of continuous processing. In this manuscript, we use process economic modeling and Monte Carlo simulations to evaluate an integrated continuous biomanufacturing (ICB) platform and conduct risk-based valuation to generate a probabilistic range of net-present values (NPVs). For a specific ten-year product portfolio, the ICB platform reduces average cost by 55% compared to conventional batch processing, considering both capital and operating expenses. The model predicts that these savings can further increase by an additional 25% in situations with higher-than-expected product demand showing the upward potential of the ICB platform. The ICB platform achieves these savings and corresponding flexibility mainly due to process intensification in both upstream and downstream unit operations. This study demonstrates the promise of continuous bioprocessing while also establishing a novel framework to quantify financial benefits of other platform process technologies.

Integrated continuous production of recombinant therapeutic proteins.

In the current environment of diverse product pipelines, rapidly fluctuating market demands and growing competition from biosimilars, biotechnology companies are increasingly driven to develop innovative solutions for highly flexible and cost-effective manufacturing. To address these challenging demands, integrated continuous processing, comprised of high-density perfusion cell culture and a directly coupled continuous capture step, can be used as a universal biomanufacturing platform. This study reports the first successful demonstration of the integration of a perfusion bioreactor and a four-column periodic counter-current chromatography (PCC) system for the continuous capture of candidate protein therapeutics. Two examples are presented: (1) a monoclonal antibody (model of a stable protein) and (2) a recombinant human enzyme (model of a highly complex, less stable protein). In both cases, high-density perfusion CHO cell cultures were operated at a quasi-steady state of 50-60 × 10(6) cells/mL for more than 60 days, achieving volumetric productivities much higher than current perfusion or fed-batch processes. The directly integrated and automated PCC system ran uninterrupted for 30 days without indications of time-based performance decline. The product quality observed for the continuous capture process was comparable to that for a batch-column operation. Furthermore, the integration of perfusion cell culture and PCC led to a dramatic decrease in the equipment footprint and elimination of several non-value-added unit operations, such as clarification and intermediate hold steps. These findings demonstrate the potential of integrated continuous bioprocessing as a universal platform for the manufacture of various kinds of therapeutic proteins.