Informa Life Sciences is part of the Knowledge and Networking Division of Informa PLC
This site is operated by a business or businesses owned by Informa PLC and all copyright resides with them. Informa PLC's registered office is 5 Howick Place, London SW1P 1WG. Registered in England and Wales. Number 3099067.
KEYNOTE PRESENTATION: Building Value Through External Stakeholder Engagement
Historically, the alkali stability of Protein A chromatography resins has lagged behind other resins used in downstream purification of antibody therapeutics. This fact, combined with the high nutrient load to which Protein A resins are exposed, introduces a larger risk of column contamination when compared with secondary and tertiary chromatographic operations. In this presentation, we present a newly designed protein A resin which offers alkali stability on par with ion exchange and hydrophobic interaction chromatography resins. Data on binding capacity, caustic stability and resin cleanability will be presented.
Recovery & Purification: Process Modeling/Streamlining Process Characterization and Process Validation
This talk will highlight the use of a multimodal cation exchange (MMCEX) chromatography column as a polishing step to reduce host cell protein (HCP) and high molecular weight species in a non-IgG production process. The reduction of impurities was achieved by influencing the effects of charge and hydrophobic interactions of the MMCEX chromatography. Specifically, buffer and protein load compositions were studied for its capacity to exploit both CEX and HIC components of the resin ligand during product elution. A high throughput robotic screen was first performed to identify key process parameters and minimize the number of lab scale chromatography experiments. Ultimately, the MMCEX column was implemented as the final chromatography unit operation in the non-IgG purification process to reduce HCP levels approximately 10-fold while maintaining process manufacturab ility an d desired product quality.
Cation exchange chromatography (CEX) operated in bind-and-elute mode is a common polishing step for removal of aggregates in purification processes of biopharmaceuticals. Depending upon the resolution of the product and the aggregates, the operating ranges for process parameters providing the acceptable clearance can be narrow. This requires tight manufacturing process controls to ensure consistent product quality and process performance. This presentation will show a case study using a monoclonal antibody with cation exchange chromatography operated in flow-through mode, resulting in a more “manufacturing-friendly” purification approach and robust alternative. The flow-through CEX was able to provide high aggregate clearance and step yield, comparable to or better than the bind-and-elute CEX method, over wider ranges of process parameters and with fewer critical process parameters for the control strategy of this step. The presentation will also illustrate advantages of flow-through CEX on manufacturing throughput, raw material consumption, and improved manufacturing process fit over bind-and-elute CEX. Data on process robustness of the flow-through CEX to resin variability versus the bind-elute CEX mode of operation will also be included.
Process characterization studies are a key component of a Quality by Design (QBD) approach, enabling improved process understanding, determination of critical process parameters (CPP), and definition of the process design space, ensuring that the process is capable of reproducibly generating safe and efficacious product for patients. While a classical design of experiments (DOE) approach is typically used for these studies, there are new statistical and data analytical techniques available that can improve experimental efficiency and better utilize the wealth of data generated from these studies. We demonstrate the potential for these new techniques with a case study of the downstream process characterization of a recombinant enzyme process. Of the four chromatography operations studied, one proved to be a particular challenge. During the initial multivariate screeni ng study, fully 25% of the experiments resulted in extremely low product recovery, indicating that the original process set-point was not robust. Because a new type of screening DOE, a definitive screening design (DSD), was used for this study, we were able to quickly and efficiently identify a new region of the design space for further characterization. Additional experiments were added in this region to support a response surface model, enabling the use of process simulation to identify a robust operating set-point and process design space. Utilization of these new statistical methods greatly improved the efficiency of the characterization of this complex chromatography operation. These techniques, especially the application process simulation for robust process engineering, can become a part of the standard toolbox for future QBD studies.
Cell Culture & Upstream Processing: Cell Culture Development
The adaptability and utility of CHO cell factories derives from exploitation of their acquired genetic/functional hyper-variation using high-throughput functional screening and selection processes which enable industry to identify, isolate and maintain cell lineages with unusual and desirable manufacturing properties. However, the propensity of clonally derived cell populations to unpredictably change their functional performance by progressive genetic drift during subculture is also a significant problem. Clonal variation is both a blessing and a curse. How can we maximize benefit from clonal variation whilst limiting its deleterious consequences – minimizing resource expenditure on stability testing? For example, our data reveal that for the vast majority of isolated CHO subclonal populations, metabolic phenotype and culture performance drift markedly over extended subculture, with only a small proportion of subclones exhibiting both high culture performance and functional stability. New methods to both unlock the synthetic potential of CHO cells and ensure these remain stably inherited are required. In this presentation I will discuss new methodologies to (i) create functionally superior “biomass intensive” clonal variants through directed evolution and (ii) measure and predict clone-specific functional stability using rapid, multiparallel chemical stress fingerprinting.
Poloxamer 188, a common surfactant utilized in biopharmaceutical manufacturing, has undergone intense scrutiny in recent years due to lot variability impacts throughout the industry. Investigations into the cause of variability have resulted in an improved understanding of poloxamers, including surfactant-cell interactions within cell culture. The generation of highly productive mammalian cell lines for biopharmaceuticals is essential to enabling a high-yield manufacturing process, and there are several steps within the cell line development process that can utilize medium containing poloxamers. DNA transfection via lipofection or electroporation methods involves delivery of DNA across the cell membrane and integration of that DNA into a host cell genome to express recombinant proteins. Cell culture media or formulated buffers have been optimized to enable effic ient gen e transfer; however, the effect of poloxamers in these solutions has not been extensively studied. We have investigated the impacts of poloxamers on various transfection methods and subsequent culture recovery, and have developed optimized processes for generating highly productive cell lines.
Analytical & Quality: Change Control and In-Country Release Testing
Drug Product, Fill-Finish & Formulations: Formulation Development Strategies for Biologics
To ensure delivery of safe and efficacious drug products to patients, biologics must remain within recommended temperature range during manufacturing, storage, shipping and handling. However environmental factors, equipment failure or packaging errors, may result in a deviation to the recommended temperature. This presentation will discuss industry practices on temperature excursions handling for biologics.
Manufacturing Strategy: Biomanufacturing Strategies for Novel Modalities and Vaccines
Univercells is offering a disruptive process featuring high-density bioreactors operated in perfusion, integrated with in-line sequential-continuous purification, resulting in a drastic reduction of manufacturing cost. This intensified and integrated manufacturing process can be fully contained into an isolator able to produce any type of viral vaccine.
This presentation examines the validation process used to define robust and redundant disconnection methodologies as container closures for intermediate and drug substance dispensing platform at cold temperatures after shipping.
Thirty years ago, monoclonal antibodies were mainly research projects which were barely represented in pharmaceutical sales. Today, mAbs represent 50% of all biological sales and the pipeline of future mAbs and other antibody constructs is rich. Dr. Hale, who was involved in the development of one of the first mAbs, Campath, will look back on the dramatic success of our industry and how the developments of antibody and protein A technologies have followed a synergistic path to bring the industry to its current state.
For >70 years Mitsubishi Chemical has been manufacturing industrial separation media for peptides and small to large molecules. Our newest media, MabSpeed™(protein A) and ChromSpeed™ (ion exchange) are specially designed for purification of biopharmaceuticals (antibody-drugs, protein-drugs, etc.) as well as large oligonucleotides for high throughput processing. The details of these products will be introduced during the seminar.
Protein A and Protein L chromatography resins are used as a capture step for intact mAbs, antibody fusion proteins and mAb fragments. Because of their unique specificity, these resins are capable of adequately removing most process impurities. They also exhibit the highest binding capacity, the widest range of mAb subclass specificity and are resistant to alkaline cleaning.
The radio-frequency impedance method for real time detection of viable biomass is established in biopharmaceutical applications. This method provides real time information of live biomass and is used extensively to monitor processes and make process decisions. This presentation covers the principle behind the impedance method, its benefits, and applications of the technology across processes, making it a robust PAT tool.
The presentation panel discusses the three perspectives (bioprocessing company, system integrator, component manufacturer) of validation methodology for single-use systems as the bioprocessing industry is confronted with producing safe therapies and vaccines while meeting government regulations and constant pressure to lower cost per dose. It takes a look at the challenges in implementation and adoption of validation methodology and the benefits of collaboration.
Exhibit Hall Theater Presentations
Alex de Winter, Managing Director, GE Ventures
Alex Lash, National, Biotechnology Editor, Xconomy
Luke Timmerman, Founder and Editor, Timmerman Report
Rebecca Robbins, Business Reporter, STAT News
Recovery & Purification: Overcoming Bottlenecks from High Cell Densities and Process Intensification
The reduction of antibody interchain disulfide bonds in harvested cultures of Chinese Hamster Ovary (CHO) cells has been reported in recent years. Antibody reduction has been attributed to high shear unit operations during cell culture recovery, causing cell lysis and the release of reducing enzymes into the clarified harvest. In the presented study, we identified disposable depth filtration as a high shear recovery unit operation that is responsible for antibody reduction in clarified cell cultures. Depth filtration process parameters that are critical to control antibody reduction, as well as quality attributes of the clarified harvest to monitor during filtration development will be discussed. Additionally, proposed risk mitigation strategies will be covered to control product quality when considering disposable depth filtration for the recovery of antibodies from CH O cell culture.
Cell Culture & Upstream Processing: Combining High Throughput Cell Culture to High Throughput Analytics
We present use cases demonstrating the automation of the development and assessment of mammalian cell lines and related upstream processes in high throughput, including seeding, selection, passaging, cryo-conservation, as well as batch and fed-batch processing and data analysis. We show how the full history of all clones can be tracked - from initial transfection all the way to their evaluation in bioreactor runs - and how this information can be combined with product quality and analytics data. These automation approaches include the integration with instruments, such as pipetting robots, colony pickers, bioreactors and bioanalyzers and deal with cell line and process development for both antibodies (IgGs, novel formats) as well as therapeutic proteins (e.g., fusion proteins).
With the potential of following different paths to market, there is interest in decreasing the time taken to complete process characterization studies. There is also a desire to improve the quality of the data obtained from them. As an example, the introduction of high throughput technologies can increase the throughput, improve experimental design and increase flexibility in respect of these aims. In this presentation we will discuss Lonza‘s strategy for upstream process characterization studies.
The presentation focuses on outlining a development approach which covers the full development cycle in upstream, from supporting early-stage clone selection through comprehensive process development leading to cGMP manufacturing to process characterization studies using high-throughput bioreactor scale-down models.
Analytical & Quality: Strategies for Continuous Improvement and Quality Control
This challenges decades of perceived wisdom on Deviation Management. The BPOG redesign of the Deviation Management System is driven by the recognition that different types of non-conformances are best solved using different investigational techniques. Minor, or low risk "events" are generally symptoms of a larger systemic process issues. Rather than conducting a comprehensive root cause analysis on each individual instance these are more effectively addressed by trending for patterns and then utilizing Continuous Improvement methodologies to address the "common cause" issues which lead to the events. More significant issues, deviations, are more likely the result of "special causes" and are effectively investigated using root cause analysis tools on an individual basis. We will explain the BPOG risk based Deviation Management Model and the case for change; show how the model works in practice and the major benefits companies have achieved by adopting it; and describe a best practice implementation model forged from experience. We will distinguish fact and fiction in the typical reasons cited for keeping the DMS model unchanged. In this session participants will learn about track and trending for low risk deviations, will review applied tools, and health authority reactions.
Errors are a part of life. With human errors accounting for approximately 50% of quality deviations and related problems within the BioPharma industry, the vital importance and real-world value of integrating human performance principles is obvious. The Lonza Error Prevention System (EPS) is a cultural change program build on proven approaches around coaching by leaders at the shop floor and error proofing of Work Instructions, Batch Records, Standard Operating Procedures as well as practices for structuring, conducting, and documenting training to assure operator competence. These practices shift the “training for compliance” paradigm to “training for competence” in GMP environment. Results have been significant - so far Human Error-related quality deviations and non-conformities have reduced by more than 50% across 12 sites globally and will continue to fall.
Therapeutic proteins are highly sensitive to manufacturing, handling and storage condition due to large size and more complex structure. Therefore, quality assessments of therapeutic proteins in real-time and continuously is of the most importance for rapid making-decision concerning the manufacturing process. However, bench-type conventional analytical tools, which have been used in traditional biomanufacturing, have restrictions to meet quality assurance requirements of current and future biomanufacturing systems. In this context, we developed a novel nanofluidic device enabling continuous monitoring of therapeutic proteins purity and bioactivity with high detection sensitivity, resolution and analysis speed. Periodic and angled nanofilter arrays served as molecular sieving structure was fabricated to achieve continuous-flow size-based analysis of therapeutic proteins in relatively simple operation, allowing for real-time analysis. In addition, nanofilter array can be designed to conduct various activity (binding) analyses for therapeutic proteins (analysis time: 30 min, limit of detection: 50 ng/mL (purity assessment) and 2 ug/mL (activity assessment)). Therefore, the device can provide significantly enhanced sensitivity, speed, and automation for therapeutic proteins quality assessment well beyond what is possible with conventional techniques, starting from manufacturing to patient administration. For demonstration of continuous monitoring of purity and bioactivity of biologic drugs using the nanofluidic PAT device, we performed a lab-scale perfusion culture of IgG1-producing Chinese Hamster Ovary (CHO) cells using a microfluidic cell retention device. The clarified harvest solution was continuously flowed into the nanofluidic device, and purity and bioactivity of IgG1 were assessed in real-time.
Manufacturing Strategy: Best Practices for Successful Technology Transfers in Biomanufacturing
Technology transfer is important preparation for clinical or commercial production of pharmaceuticals, especially when production will be executed at external manufacturing sites. While technology transfer focuses on transferring process and product knowledge, site readiness tasks, including those for quality and supply requirements, are of the same significance. Effective integration of tech transfer and site readiness will ensure success of production campaigns. This presentation will address details of site readiness tasks and critical documents, and discuss how to manage and integrate site readiness with technology transfer. Examples of incidents and lessons learned will also be presented.
Technology transfer leading to a successful validation of a Monoclonal Antibody process at a CMO site is a complex task that would need seamless interaction between various functions of the sending and receiving organizations. The success of the process is very much dependent on the technical depth, personal trust and the strength of the relationships established between the team members across the various functions in both organizations. The interactions begin with the due diligence process and builds through laboratory scale technology transfer leading to engineering/GMP campaigns and ends with a successful validation. Depending on the stage in development of the molecule being technology transferred, the sending organizations could have personnel involved from both development and manufacturing making the process more complex. The talk will outline the role played by the various functions starting from Technical Services to Quality Assurance and will include functions like Shipping and Supply Chain.
Drug Product, Fill-Finish & Formulations: Formulation Development Strategies for Biologics
High-concentration biologic formulations are challenging to develop due to a dramatic increase in solution viscosity at elevated protein concentration. The large viscosity is driven by attractive protein-protein interactions (PPIs), which promote long-lived clusters and networks in solution. Several formulation strategies exist to mitigate these attractive PPIs including adjusting the solution pH, ionic strength, and adding arginine. However, these strategies are limited and do not work for every molecule. Unfortunately, alternative solutions are lacking. Here we will disclose a novel excipient that lowers the viscosity of high-concentration biologics formulations. The decrease in viscosity is associated with a decrease in attractive PPIs, as measured by dynamic light scattering. Rigid-body docking simulations indicate that the excipient preferentially binds positively charged residues, such as arginine, and aromatic hydrophobic residues, such as tyrosine and phenylalanine, suggesting the dominant interaction mechanisms are π-cation interactions and π-π stacking interactions, respectively. Moreover, we show that typical additives, such as NaCl and arginine are ineffective in lowering the solution viscosity. Most importantly, the toxicology and pharmacokinetics/dynamics of the excipient are well-established, facilitating its use in biologics formulations from a regulatory stand point.
Clinical use of biologics often requires further manipulation of drug product, including transfer and dilution steps to ensure that the correct dose can be administered. Formulation scientists are an important partner of clinical study teams to address potential challenges arising e.g. through contact of dilute solutions with infusion bags, lines and filters. In-use stability and simulated administration studies are performed to understand and mitigate potential risks.