Development towards a high-titer fed-batch CHO platform process yielding product titers \u3e 10 g/L

Roche’s current Global Cell Culture Platform (GCCP) using chemically-defined media was implemented in 2012 and has been successfully used in process development and clinical manufacturing for numerous molecules. Several minor version changes have been implemented since its inception mainly to further optimize product quality requirements. However, high lactate levels have been observed in several projects using our CHO-K1-M GS host cell line (Random Integration host), resulting in sub-optimal culture performance when not addressed by process modifications (e.g., off-platform pH changes, etc.). Understanding the “triggers” for undesirable lactate metabolism and identifying levers to control lactate metabolism are keys in improving process robustness and enabling further advances in platform process optimization towards higher titers required for high-demand products. Using a lactogenic model cell line, we examined numerous potential lactate levers including starting osmolality of production media and other factors that can mitigate the buildup of in-process osmolality (e.g., media components, media powder concentration, feed strategies, and process parameters). The results from these studies were then used to further optimize our existing platform media and process to develop a high titer proof-of-concept fed-batch process yielding \u3e 10 g/L. We also investigated the optimization of media solubility and stability of our proprietary liquid media, thus enabling the development of new highly concentrated liquid media which are required for high titer processes. Case studies that demonstrate the applicability of the newly developed high titer process with numerous mAb producing cell lines including our new Targeted Integration host will be discussed. The optimized “traditional” fed-batch process may ultimately lead to our next generation platform process, which still fits within our current manufacturing network, but will significantly reduce cost of goods and runs required to support clinical and commercial production of our biopharmaceutical proteins

Implementation activities for a chemically-defined media platform to minimize media variability impact to cell culture performance and product quality

Chemically-defined media have been developed for CHO cell culture platform processes. During implementation of the media at multiple scales and sites globally, additional activities were required to seamlessly balance practical considerations with desired process and product quality performance. Three main focus areas were further investigated and optimized to enable the platform implementation: 1) media preparation, filtration, and post-filtration stability; 2) media and supplement viral barrier compatibility; and 3) and raw material variability considerations. Overall, the effort provided an outlined approach for media implementation in the global network including scale and site-specific case studies where novel options were required. The data generated supported successful implementation of chemically-defined platform media at different global sites within our network. Manufacturing-scale media preparations met acceptance criteria and resulted in comparable cell culture performance and product quality to controls. Practical challenges were identified and risk mitigation efforts initiated. The challenges were primarily related to poorly-defined media preparation mixing environments as a result of variable equipment availability and use. Filtration capacity acceptance criteria were met in the scale-down model and large-scale filtration events were successful. HTST-compatibility studies showed that HTST-treated media performed comparably to control media in cell culture performance and product quality. Post-filtration media stability investigations led to the identification of key risk factors including temperature and agitation variation during storage, nucleation and media destabilization as a result of specific handling practices, and the impact of specific process-contacting materials. Options for mitigating the media stability risks and enabling successful implementation were developed and showed no negative impact for the application. Finally, platform media modifications were developed to mitigate raw material variation and product quality challenges. The overall evolution of the media platform to ensure flexibility and successful implementation will be summarized

The Effect of Adherence to Dietary Tracking on Weight Loss: Using HLM to Model Weight Loss over Time

The role of dietary tracking on weight loss remains unexplored despite being part of multiple diabetes and weight management programs. Hence, participants of the Diabetes Prevention and Management (DPM) program (12 months, 22 sessions) tracked their food intake for the duration of the study. A scatterplot of days tracked versus total weight loss revealed a nonlinear relationship. Hence, the number of possible tracking days was divided to create the 3 groups of participants: rare trackers (\u3c33% total days tracked), inconsistent trackers (33–66% total days tracked), and consistent trackers (\u3e66% total days tracked). After controlling for initial body mass index, hemoglobin A1c, and gender, only consistent trackers had significant weight loss (−9.99 pounds), following a linear relationship with consistent loss throughout the year. In addition, the weight loss trend for the rare and inconsistent trackers followed a nonlinear path, with the holidays slowing weight loss and the onset of summer increasing weight loss. These results show the importance of frequent dietary tracking for consistent long-term weight loss success

Addressing Diabetes Distress in Self-Management Programs: Results of a Randomized Feasibility Study

Background: West Virginia ranks 1st nationally in the prevalence of hypertension (HTN; 43.8%) and diabetes (16.2%). Patients with type 2 diabetes mellitus (T2DM) are distressed over physical and psychological burden of disease self-management. Methods: This study investigated the effectiveness of an intervention to reduce diabetes distress and outcomes [glycemic control, blood pressure (BP)] among T2DM adults with comorbid HTN. Participants were randomized to a 12-week diabetes and hypertension self-management program versus a 3-month wait-listed control group. Trained health coaches and experts implemented the lifestyle program in a faith-based setting using an adapted evidence-based curriculum. Twenty adults with T2DM and HTN (n=10 per group) completed baseline and 12-week assessments. Diabetes distress was measured by using a validated Diabetes Distress Survey (17-item Likert scale; four sub-scales of emotional burden, physician related burden, regimen related burden, and interpersonal distress). Baseline and post-intervention changes in diabetes distress were compared for both groups; reduction in distress in the intervention groups are depicted using waterfall plots. The mean age, HbA1c and BMI were 55 ± 9.6 years, 7.8 ± 2.24 and 36.4 ± 8.8, respectively. Diabetes distress (total; mean) was 1.84±0.71. Results: Participants reported higher diabetes distress related to emotional burden (2.1±0.94) and regimen-related distress (2.0 ± 0.74); physician-related distress was the lowest (1.18±0.64). In general, diabetes distress reduced among intervention participants and was especially significant among those with HbA1c ≤ 8% (r=0.28, p=0.4), and systolic/diastolic BP ≤140/80 mm Hg (r=0.045, P=0.18). Implications: Findings suggest that lifestyle self-management programs have the potential to reduce diabetes distress

The relationship between children's physical activity and family income in rural settings: A cross-sectional study

AbstractObjectiveTo examine potential differences in children's physical activity and parent support of their children's physical activity based on family income within the rural setting.MethodsA cross-sectional survey of 566 parents of children (5–15years-old; mean=7.7years; standard deviation=2.4) living in rural West Virginia from 2010 to 2011 was conducted. Children were recruited and had participated in a school-based health screening program.ResultsOverall, parents from a rural setting reported that their children engaged in an average of five days of physical activity for at least 60min. Upon closer examination, children from lower-income families engaged in more physical activity, on average, than children from higher income families per parent report (mean=6.6days, confidence interval 95%=4.9–6.0 vs. middle-income mean=5.0, confidence interval 95%=4.4–5.3 and highest-income mean=4.5, confidence interval 95%=4.1–4.7; p=.01). Rural parents supported their children's physical activity in numerous ways. Parents with the lowest incomes were more likely than parents from higher income families to encourage their children to be active and use their immediate environment for play and to be directly involved in physical activity with their children. More affluent parents were more likely to transport their children to other activity opportunities than parents from the lower income brackets.ConclusionsLower income families may utilize their immediate environment and encourage activity among their children whereas more affluent families focus on organized opportunity more often than lower income families. These findings emphasize the need to conceptualize the role family income plays in physical activity patterns and the potential benefit it provides to some families

An Evaluability Assessment of the West Virginia Physical Activity Plan, 2015: Lessons Learned for Other State Physical Activity Plans

Background The US National Physical Activity Plan (NPAP) was released in 2009 as a national strategic plan to increase physical activity (PA). The NPAP emphasized implementing state and local PA pro- grams. Dissemination of information about NPAP has been lim- ited, however. Community Context West Virginia is a predominantly rural state with high rates of chronic diseases associated with physical inactivity. In 2015 an evaluability assessment (EA) of the West Virginia Physical Activ- ity Plan (WVPAP) was conducted, and community stakeholders were invited to participate in updating the plan. Methods A good EA seeks stakeholder input, assists in identifying program areas that need improvement, and ensures that a full evaluation will produce useful information. Data for this EA were collected via national stakeholder interviews, document reviews, discussions among workgroups consisting of state and local stakehold- ers, and surveys to determine how well the WVPAP had been im- plemented. Outcome The EA highlighted the need for WVPAP leaders to 1) establish a specific entity to implement local PA plans, 2) create sector-spe- cific logic models to simplify the WVPAP for local stakeholders, 3) evaluate the PA plan’s implementation frequently from the out- set, 4) use quick and efficient engagement techniques with stake- holders when working with them to select strategies, tactics, and measurable outcomes, and 5) understand the elements necessary to implement, manage, and evaluate a good PA plan. Interpretation An EA process is recommended for other leaders of PA plans. Our project highlights the stakeholders’ desire to simplify the WVPAP so that it can be set up as a locally driven process that engages communities in implementation

The Importance of Utilizing Diabetes Prevention Programs

Quality Insights and guests presented an innovative look at how the National Diabetes Prevention Programs are utilized and tracked throughout Pennsylvania and West Virginia. This webinar will bring awareness to healthcare providers, partners, and stakeholders about the recommendations on screening, testing and referring to National Diabetes Prevention Programs

The Majorana Neutrinoless Double-Beta Decay Experiment

The proposed Majorana double-beta decay experiment is based on an array of segmented intrinsic Ge detectors with a total mass of 500 kg of Ge isotopically enriched to 86% in 76Ge. A discussion is given of background reduction by: material selection, detector segmentation, pulse shape analysis, and electro-formation of copper parts and granularity. Predictions of the experimental sensitivity are given. For an experimental running time of 10 years over the construction and operation of Majorana, a half-life sensitivity of ~4x10^27 y (neutrinoless) is predicted. This corresponds to an effective Majorana mass of the electron neutrino of ~0.03-0.04 eV, according to recent QRPA and RQRPA matrix element calculations.Comment: 10 pages, 7 figure