Biomedical Engineering and Bioengineering Commons^™

Cder's Emerging Technology Team, Kurt Brorson, Sau Lee

Integrated Continuous Biomanufacturing II

The newly formed Emerging Technology Team (ETT) draws membership from all CDER quality review, research and inspection functions, including Office of Biotechnology Products. The goal of the ETT is provide a primary point of contact for external inquiries regarding emerging technology in pharmaceutical and biotechnology manufacturing and quality control. The ETT will partner with review offices in a cross-functional manner and identify regulatory strategy and resolve roadblocks to new technologies relating to existing guidance, policy or practices related to review or inspection. The ETT’s initial focus will be innovation on novel products, manufacturing processes, or testing technologies or processes to …

Monitoring And Control Of Reproducibility In Quasi-Continuous Integrated Production Processes Of Active Pharmaceutical Ingredients, Reiner Luttman, Sven Oliver Borchert, Jan Patrick Voss, Gesine Cornelissen

Integrated Continuous Biomanufacturing II

The development of integrated production processes include the combination and transformation of current batch oriented unit operations into linked sequential/parallel production strategies.

The presented process starts with a two-stage upstream consisting of cell cultivation and subsequent protein production, which in turn results in a five step downstream process, consisting of cell clarification via a separator, retention of cellular debris using microfiltration, concentration of the secreted product by ultrafiltration with subsequent buffer exchange through diafiltration, followed by a final purification using column chromatography.

The three main operations cell breeding, protein production and the complete downstream line are running in series, but …

Exploring Continuous And Integrated Strategies For The Up- And Downstream Processing Of Human Mesenchymal Stem Cells, Barbara Cunha, Tiago Aguiar, Ricardo Silva, Cristina Peixoto, Manuel Carrondo, Margarida Serra, Paula Alves

Integrated Continuous Biomanufacturing II

The integration of up- and downstream unit operations can result in the elimination of hold steps, thus decreasing the footprint, and ultimately can create robust closed system operations. This type of design is desirable for the bioprocess of human mesenchymal stem cells (hMSC), where high numbers of pure cells, at low volumes, need to be delivered for therapy applications. The aim of this work is to perform a proof of concept of the integration of a continuous perfusion culture in bioreactors with a tangential flow filtration (TFF) system for the concentration and washing of hMSC. In particular, we have evaluated …

Pat Concepts For Chromatography: Real-Time Monitoring, Real-Time Control, And Cause Of Error Diagnostics, Nina Brestrich, Tobias Hahn, Adrian Sanden, Till Briskot, Jurgen Hubbuch

Integrated Continuous Biomanufacturing II

In the recent years, the growing competition in the biopharmaceutical industry due to biosimilars combined with market fluctuations and increasing product pipelines have given rise to the evaluation of continuous processing. Advantages include smaller equipment that may be disposable, an overall process intensification as well as steady-state operation. As variability should be managed by the continuous process itself to insure a consistent product quality, the development of advanced Process Analytical Technology (PAT) concepts is one crucial aspect for the implementation of continuous processing.

In chromatographic protein purification, process variability can lead to variations in retention volumes or peak shapes of …

Quality Systems For Continuous Manufacturing, Ron Branning

Integrated Continuous Biomanufacturing II

Continuous manufacturing is a familiar concept in large volume commercial commodity operations such as glass and plastic production. In the wake of the Tylenol cyanide poison scare in the 1970’s, pharmaceutical production at the J&J subsidiary McNeil converted to virtually closed, continuous manufacturing. Several Biotech firms tried to develop fermentation unit operations in the 1990’s that were based on the continuous manufacturing concept. Today there are a number of initiatives to update and realize the concept from bench top, such as MIT and Novartis’ sponsorship of start up firm Continuus’ production of solid dosage forms in Boston, to biodefense tractor-trailer …

Dynamic Oncolytic Measles Virus Production, Tanja Grein, Denise Salzig, Michael Muehlebach, Peter Czermak

Integrated Continuous Biomanufacturing II

Oncolytic viruses can be effective weapons against cancer with few treatment options. For example the tissue culture–adapted Edmonston strains of measles virus (MV) have altered its receptor specificity and became selectively oncolytic with attenuated pathogenicity. Russel et al. showed in 2014 full remission in an advanced stage multiple myeloma patient after systemic application of genetically modified MV. In this clinical trial, the patient was treated by intravenous infusion of 10¹¹ TCID₅₀ (50% tissue culture infectious dose) - of an engineered MV encoding human sodium iodide symporter. Appropriate medical treatment with oncolytic viruses calls for high concentrations and absolute …

Transforming The Present Into The Future With Uncertainty And Imprecision, Chales Cooney

Integrated Continuous Biomanufacturing II

No abstract provided.

How Continuous-Like Processes Improve Affordability For Viral Vaccines - Example Of Laiv, Jose Castillo

Integrated Continuous Biomanufacturing II

Designing a continuous process for a life viral vaccine could be seen useless, and challenging, as the virus infection will kill the cell substrate after 2-5 days of infection.

From the technical perspective, chaining the operations of cell culture under perfusion and viral production, clarification and capture / concentration is an efficient option to setup a small scale continuous automated process to manufacture massive amounts of viral vaccines.

We will show how beneficial it can be in terms of process development, manufacturing simplicity, capital expenditure and cost of production. Also, we will show how this approach can help producers of …

Advanced Computational Tools To Enhance Continuous Monoclonal Antibody Production, Maria M. Papathanasiou, Ana Quiroga, Athanasios Mantalaris, Efstratios Pistikopoulos

Integrated Continuous Biomanufacturing II

Leading pharmaceutical companies invest high percentage of their revenue in the improvement of existing technologies used for the production of monoclonal antibodies (mAbs). Recently, there has been a paradigm shift towards the development of continuous/quasi-continuous purification operations, aiming to reduce capital and operational costs [1]. At the moment, however, there are no standardized methods and/or tools that can be used for global control and monitoring of integrated processes.

Mathematical models and advanced computational tools can be the key for the development of robust, integrated processes, as they can provide valuable insight in the process dynamics and ensure optimal operation [2]. …

Protein Refinery Operations Lab (Pro Lab): A Sandbox For Continuous Protein Production & Advanced Process Control, Mark Brower, David Pollard, Finn Hung

Integrated Continuous Biomanufacturing II

Significant strides towards implementation of continuous bioprocessing are being made at an ever increasing rate. Advances in technology for traditional unit operations such as cell-retention devices in perfusion cell culture, continuous multi-column chromatography (CMCC) and single-pass tangential flow filtration have led to demonstrations of both semi-continuous and fully-continuous protein production processes operating at periodic steady states at the pilot-scale. Previous proof of concept work at Merck & Co., Inc. has shown an automated (DeltaV) and single-use monoclonal antibody (mAb) purification scheme through Protein A CMCC and pH viral inactivation with minimal human interaction for 30 days fed from a perfusion …

A Regulatory Perspective On Continuous Perfusion Production Of Rfviii, Robert Kozak

Integrated Continuous Biomanufacturing II

Continuous perfusion production has enabled flexible manufacturing of rFVIII, a large complex Biotech Product, for over two decades. Continuous improvement has driven frequent process, equipment and facility changes successfully performed utilizing comparability exercises.

Challenges in evaluating changes made to a continuous perfusion process includes assessment of impact to quality product attributes throughout the entire fermentation campaign which can be months in duration. Examples of process changes requiring more studies, those successfully supported by small scale development runs and API commercial characterization and an example resulting in non-implementation will be reviewed. Impact of changing Regulatory Environment on submission package requests will …

Small-Scale Platform For Rapid On-Demand Manufacturing Of Recombinant Proteins, J. Christopher Love

Integrated Continuous Biomanufacturing II

The current timeline for manufacturing high-quality recombinant proteins used for therapeutics and vaccines, and delivering them to patients, typically requires many months. There are many circumstances, however, where the rapid production, release, and delivery of these biologics could address unmet medical needs, including shortages, emergency situations, and pandemics. This talk will present a new platform developed to produce biologic medicines on demand called Integrated and Scalable Cyto-Technology (InSCyT). This platform comprises an integrated, subliter-scale portable system for the (semi)continuous operation of fermentation, filtration of cell debris from secreted product, innovative affinity-based purification, polishing, and finishing. This program also emphasizes integrated …

Regulatory Challenges Of Continuous Biomanufacturing, Andy Papas

Integrated Continuous Biomanufacturing II

The interaction with regulatory agencies on the biomanufacturing of therapeutic proteins, mAbs, or other large molecules occurs throughout the drug development of the biologic; it begins with the original IND, accelerates at the licensing phase and continues throughout the product’s life cycle. There are many regulatory challenges along this journey.

Because of uniqueness of continuous biomanufacturing, one challenge that occurs throughout the product’s lifecycle is the inevitable change in Agency product reviewers who many times need to be educated on the nuances of your process or have a different opinion or expectation on how things should be done. As such, …

2d Cine Dense With Low Encoding Frequencies Accurately Quantifies Cardiac Mechanics With Improved Image Characteristics, Gregory J. Wehner, Jonathan D. Grabau, Jonathan D. Suever, Christopher M. Haggerty, Linyuan Jing, David Powell, Sean M. Hamlet, Moriel H. Vandsburger, Xiaodong Zhong, Brandon K. Fornwalt

Biomedical Engineering Faculty Publications

BACKGROUND: Displacement Encoding with Stimulated Echoes (DENSE) encodes displacement into the phase of the magnetic resonance signal. The encoding frequency (k_e) maps the measured phase to tissue displacement while the strength of the encoding gradients affects image quality. 2D cine DENSE studies have used a k_e of 0.10 cycles/mm, which is high enough to remove an artifact-generating echo from k-space, provide high sensitivity to tissue displacements, and dephase the blood pool. However, through-plane dephasing can remove the unwanted echo and dephase the blood pool without relying on high k_e. Additionally, the high sensitivity comes …

Design And Control Of Chromatography Step In An Integrated Column Sequence, Bernt Nilsson, Niklas Andersson, Anders Holmqvist, Anton Sellberg, Peter Tiainen, Arne Staby

Integrated Continuous Biomanufacturing II

Integration of the purification steps in downstream processing is one way to minimize the number of storage tanks and reduce hold-up time. The result is an integrated column sequence that performs straight through processing of the target protein, with minimal time from expression to formulation. This paper discusses the overall integration design and its expected performance, together with a more detailed analysis of the step behavior based on column simulations. The optimal design of the individual steps in the sequence is presented, both with robustness margins and local control system, as well as with and without local recycle.

An optimization …

Integrated Continuous Processing For The Manufacture Of Monoclonal Antibodies, Daniel Karst, Fabian Steinenbach, Massimo Morbidelli

Integrated Continuous Biomanufacturing II

Continuous manufacturing is currently being considered by the Biopharmaceutical Industry not only for the classical reasons which make continuous operation preferred over the batch one, but also for recent initiatives of the regulatory agencies. We discuss here a series of experiments where a perfusion reactor with CHO cells for the production of a monoclonal antibody has been operated in the continuous mode and connected to a two column continuous protein A chromatographic unit for product capture. A few steady states are examined and the use of simulation models for process design and control is illustrated.

Integrated Continuous Bioprocessing – Opportunities And Challenges, Mats Akesson, Martin Heitmann, Ditte Skipstrup, Peter Tiainen

Integrated Continuous Biomanufacturing II

We have developed an integrated continuous processing framework for end-to-end production of complex fragile proteins based on perfusion cultivation and automated multi-step purification [1,2,3]. Upstream, the integrated system consists of a stirred tank bioreactor with an ATF cell retention system. The clarified harvest directly enters an off-the-shelf ÄKTA chromatography system converted into a continuous purification unit. Two alternating capture columns precede an automated multi-step purification train with full flexibility and control of individual columns.

The integrated set-up enables compact automated bench-top factories converting cell culture media to purified protein in an efficient manner without intermediate storage. It provides monitoring of …

Continuous Culture And Downstream Processing Of Algae With Recycle: An Integrated Large-Scale Approach For Production Of Renewable Crude Oil, Matt Croughan

Integrated Continuous Biomanufacturing II

In order to have a significant impact on energy security and reduction of greenhouse gasses, the production of advanced biofuels must be increased to billions of gallons per year. Scale up of phototrophic algae bioprocessing for renewable oil production has been attempted for decades and has recently reached new levels of performance and scale. For over three years, Sapphire has successfully operated the world’s largest algae farm, consisting of 100 acres of ponds in New Mexico, used to feed an integrated conversion and extraction unit based on high temperature liquefaction technology. All aspects of the process are run on a …

Continuous Beer Production Methods: A Review Of Chances And Risks, Konrad Müller-Auffermann

Integrated Continuous Biomanufacturing II

Beer brewing is one of the oldest biotechnological processes applied by humans. The production methods are comparatively complex and time consuming, which is the reason why inventors and scientists have steadily attempted to improve its manufacturing.

During the industrial revolution more and more processes where done continuously in order to increase production efficiency. Breweries where trying to modify the classical batch modes into a continuous production process but were unsuccessful. Nearly all approaches failed due to changes of the beer quality.

The presentation shall give a brief overview of different continuous production concepts tested in the past 150 years. Furthermore …

Laboratory Scale Continuous Linear Purification As A Development Tool For Recombinant Blood Protein Processing, Using Chromatographic Resins And Membranes, Michael Hughson, Rimenys Carvalho, Thaynna Araujo Cruz, Leda Dos Reis Castilho

Integrated Continuous Biomanufacturing II

Continuous processing offers significant advantages for the processing of unstable recombinant products such as therapeutic plasma proteins. As such we have established a development platform to assess potential purification steps as part of a continuous linear process using a standard AKTA Explorer. Following a consistent format of IEX membrane to HIC membrane to affinity resin, we were able to rapidly investigate multiple purification pathways for a recombinant blood protein. Using membranes as the first two steps enables a 3-step purification process to be carried out in 3 to 4 hours, with a total turnaround of 5 to 6 hours including …

Continuous Countercurrent Tangential Chromatography For Purification Of High Value Therapeutic Proteins, Andrew Zydney, Oleg Shinkazh, Boris Napadensky, Amit Dutta

Integrated Continuous Biomanufacturing II

One of the main challenges in the development of integrated continuous processes for the production of high value therapeutic proteins is the replacement of traditional column chromatography, which tends to dominate current downstream processing. Continuous Countercurrent Tangential Chromatography (CCTC) has been designed to provide truly continuous product capture and purification using a column-free system that overcomes many limitations of traditional column chromatography. All operations in CCTC are conducted on a moving slurry that is continuously pumped through a cascade of static mixers (for mixing and residence time) and hollow fiber membrane modules (for separation of the fluid phase from the …

Modeling Perfusion At Small Scale Using Ambr15tm, Dustin Davis, Delia Lyons, Scott Ross

Integrated Continuous Biomanufacturing II

Reaching cell densities higher than 80 million with the minimum possible perfusion rates is a goal for an increasing proportion of processes developed by biopharmaceutical companies. With the goal of fulfilling the industry needs for better commercial and customized perfusion media, SAFC evaluated different small-scale perfusion models to achieve an efficient work flow that can accommodate perfusion systems. SAFC successfully uses an optimized work flow for the development of media and feeds for fed-batch cell culture that integrates high-throughput screening, statistical tools and bench-top bioreactor scale studies. In this model, 96-deep well plates are used for the initial high throughput …

Monitoring Intracellular Component Pools To Identify Steady State In Mammalian Cell Perfusion Culture, Daniel Karst, Robert Steinhoff, Marie Kopp, Renato Zenobi, Massimo Morbidelli, Miroslav Soos

Integrated Continuous Biomanufacturing II

Perfusion cultures of mammalian cells are a well-considered alternative in the process development for new biologic products. Stable operation of the culture should eventually lead to a steady state, positively effecting product quality. Although macroscopic variables such as viable cell density, main metabolite or product concentrations are observed constant, little is known about the extent of steady state on intracellular level.

In this study intracellular component pools of CHO cells were monitored at different steady states in perfusion cultures using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). A novel single extraction was used to separate metabolite, …

Biosc® Predict Simulation Software: Flexibility And Optimization Of Your Multi-Column Process, Fabien Rousset

Integrated Continuous Biomanufacturing II

For a few years now, the biopharma industry has started implementing continuous processing, and more specifically continuous chromatography, to improve process economics together with process control and understanding. A simulation / optimization software has been developed to benefit from modelization by accommodating batch process data to multicolumn processes with associated optimal operating costs and reduced footprint. The presentation will highlight how the flexibility of the BioSC® and its simulation software BioSC® Predict is used to optimize affinity, ion exchange and isocratic processes in order to decrease media and buffer consumption and save lab operating time.

Medium Optimization Case Study For Continuous Upstream Process, Rashmi Kshirsagar, Alan Gilbert

Integrated Continuous Biomanufacturing II

Based on other mature industries, continuous upstream process is a logical replacement for current fed batch operations. However most industrial medium development has focused on the biological requirements for fed batch and therefore focused on the needs of stationary phase production. There is not an a priori expectation that growth and stationary phase requirements are identical. Yet an ideal continuous upstream process requires some combination of both. An optimal continuous upstream process requires a high cell density similar to fed batch operations. There is also some minimum growth rate required in order to match the combined death and cell removal …

Alternating Flow Filtration As An Alternative To Internal Spin Filter Based Perfusion Process: Impact On Productivity And Product Quality, Guillermina Forno, Cristian Paillet, Laura Mauro, Ignacio Amadeo, Eduardo Orti

Integrated Continuous Biomanufacturing II

The application of an alternating flow filtration (ATF) system adapted to a stirred tank bioreactor as an alternative to internal spin filter was studied using CHO cells producing a fusion protein. As expected, it was possible to reach higher cell densities (maximum 50 x 10⁶ vs 20 x 10⁶ cells/ml) and achieve longer runs with the ATF device (50 days vs 30 days), since it is known that this cell retention device is less affected by fouling.

Even tough specific cell yield was similar for both systems; volumetric production of the reactor using the ATF was almost 70% …

A Novel Plant Cell Culture Platform For Semicontinous Production Of Recombinant Proteins: Butyrylcholinesterase As A Case Study, Jasmine Corbin, Bryce Hashimoto, Kalimuthu Karuppanan, Raymond Rodriguez, Somen Nandi, Brian Roberts, Amy Noe, Karen Mcdonald

Integrated Continuous Biomanufacturing II

In this paper we describe a novel biomanufacturing production platform that utilizes transgenic rice cell suspension cultures for efficient semicontinuous cell culture (SCC) production of recombinant proteins. The production platform utilizes a metabolically regulated promoter, a secretion signal peptide that enables secretion out of the cell for ease of recovery/purification, coupled with an efficient semicontinuous operational strategy that allows independent optimization of growth and production phases. In addition, long term operation (up several months¹) is possible by maintaining viable biomass within the bioreactor, thereby reducing the need for long seed trains, as well as minimizing turn-around time, CIP …

Integration Of Continuous Ethanol Precipitation And Flocculation Into Manufacturing Of Antibodies, Alois Jungbauer, Nikolaus Hammerschimidt, Peter Satzer

Integrated Continuous Biomanufacturing II

Precipitation and flocculation are optimal unit operations for continuous capture of proteins from culture supernatant. Precipitation and flocculation can be operated in a real continuous manner. The methods can be combined and after this capture step the precipitate can be stored as a concentrated solution or even as a precipitate. We have developed several precipitation/ flocculation protocols for capture of antibodies from culture supernatant. These include the combinations of CaCl₂ flocculation with either cold ethanol precipitation or PEG precipitation and octanoic acid precipitation with PEG precipitation. The precipitation conditions have been screened in microtiter plates or in case of …

Evaluation Of Novel Cex Resin For Continous Processing Of Mab Purification, Takuya Muramoto, Satoshi Murakami, Shinsuke Higuchi, Keiji Iwamoto

Integrated Continuous Biomanufacturing II

We have evaluated novel cation exchange resin and membrane which is potentially suited for continuous process in biologics manufacturing.

Typically cation exchange chromatography (CEX) is used in a bind/elute (B/E) mode for the MAb process since MAb act as cation at the buffer condition used for CEX due to the pI from neutral to weak basic. Generally, purpose of CEX chromatography is to remove aggregates and other impurities like HCP and DNA. Especially, aggregate removal is of interest to the industries. A continuous process in MAb downstream process can solve several bottlenecks in typical batch process. However its capability cannot …

Impact Of Closed And Continuous Processing On Biopharmaceutical Facility Layouts, Marc Pelletier

Integrated Continuous Biomanufacturing II

In response to the uncertainty of ever-changing product pipelines, the biotech industry has been driven to develop flexible manufacturing solutions. These facilities can mobilize quickly to provide a nimble response to sudden changes in production demands. The industry has concurrently improved its ability to assess and effectively mitigate risks to manufacturing processes by developing engineering solutions that isolate the bioprocess from the environment housing it. This presentation will illustrate how the implementation of continuous closed bioprocessing impacts the design of biopharmaceutical manufacturing facilities. The impact of closed continuous bioprocessing on facility size, capital cost and operating cost will also be …