Bioprocess Alert

Let us take you on a journey to the future of smart sensor technology with an excerpt from our upcoming publication, Sensing the Future. Preview the new book by downloading this free nine-page article, titled The Future of Intelligent Process Sensors, to get an inside look into where we’ve been, where we are now, and most importantly, where we’re going.
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This Special Issue of Processes entitled “Bioprocess Monitoring and Control” presents novel examples of on-line monitoring and closed loop control techniques applied to different bioprocesses. The accepted manuscripts cover a range of important topics in different bioprocess areas, where microorganisms, bird’s eggs, and humans are involved. Different techniques such as those for the construction of sensors, the production of a biocontrol agent, scaling up procedures, the application of observers, closed loop control, and the model-based monitoring of a downstream process are presented. The accepted manuscripts are nine original research papers and three reviews.
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Defining a suitable agitation strategy is essential for the successful development of upstream mammalian bioprocesses. Such a strategy can drive oxygen mass transfer within the bio- reactor system to effectively support the aerobic needs of the expanding culture. An inadequate agitation strategy can result in the emergence of chemical gradients within the system. The existence of such nonuniformity within a bioreactor can result in extracellular conditions which may be unfavorable to the overall growth of the cell culture. Hypoxic conditions in a bioreactor are defined by low dissolved oxygen concentrations.
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When technology groups add consulting services to their portfolio, some customers are initially skeptical. The inevitable question is whether a major corporation like Siemens can offer advice independently of its own products and technological solutions. Siemens Digitalization Consulting has already accompanied numerous decision-makers at pharmaceutical companies on their path to digitalization. Its team of experts with backgrounds in industry, IT, and automation enables consultants to work closely in partnership with customers to develop and define their digitalization roadmap. The roadmap is based on the customers’ existing infrastructure and landscape – IT, IoT (Internet of Things), and OT (operational technology), among others – and is tailored to their business challenges and drivers. 
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In 2019, G-CON provided the cleanroom solution for GenCure’s new multi-product cell therapy facility. The new facility, which anchors the VelocityTX innovation center in San Antonio, Texas, expands GenCure’s manufacturing capabilities, providing state-of-the-art manufacturing services to therapeutic developers. This case study captures the experience from the customer's perspective of the use of prefabricated cleanrooms for cell therapy manufacturing.
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Process control strategies based on the physiological status of cells have recently been used to enhance mammalian cell culture productivity and robustness. In this study, we investigated the feasibility of using full-spectrum dielectric spectroscopy for detecting shifts in cell physiology and as a feedback tool to increase process efficiency. Multi-frequency permittivity spectra were collected from cell culture processes in which apoptosis was induced by glucose depletion, nutrient depletion, or chemical treatment. Meanwhile, key parameters of critical frequency (fc) and Cole-Cole alpha (α) were calculated in real time from the β-dispersion curve and correlated to data from off-line viability measurements. Results show that physiological changes in apoptotic cells were reflected in the on-line parameters earlier than from off-line methods.
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Implementing a personalised feeding strategy for each individual batch of a bioprocess could significantly reduce the unnecessary costs of overfeeding the cells. This paper uses lactate measurements during the cell culture process as an indication of cell growth to adapt the feeding strategy accordingly. For this purpose, a model predictive control is used to follow this a priori determined reference trajectory of cumulative lactate.  Each experimental set-up is performed in triplicate and for each run an individualised model-based predictive control (MPC) controller is developed. All process models exhibit an accuracy of 99.80% ± 0.02%, and all simulations to reproduce each experimental run, using the data as a reference trajectory, reached their target with a 98.64% ± 0.10% accuracy on average. Learn More

Bacteriophages are bionanoparticles with several applications in different biotechnology‐based products. Among them, vaccines have the potential to treat antibiotic‐resistant bacteria and parasitic infections. Traditional methods for their recovery and purification rely on precipitation with polyethylene glycol (PEG) and NaCl. However, the applicability of such an approach is limited, due to large‐scale technical constrains. Recently, our research group developed a bacteriophage M13 recovery and purification strategy using Aqueous Two‐Phase Systems (ATPS), simplifying the methodology and, potentially, reducing costs. This work aims to develop an economic contrast between ATPS and the traditional PEG precipitation method at different operation scales (10 to 1000 L bioreactor volume) to determine the applicability of the ATPS methodology at large scale. Learn More

A process characterization is a regulatory imperative for process validation within the biopharmaceutical industry. Several individual steps must be conducted to achieve the final control strategy. For that purpose, tools from the Quality by Design (QbD) toolbox are often considered. These tools require process knowledge to conduct the associated data analysis. They include cause and effect analysis, multivariate data analysis, risk assessment and design space evaluation. Here, we present a novel workflow that shows how simple data analysis tools can be used to investigate the process holistically. This results in a significant reduction of the experimental effort and in the development of an integrated process control strategy. Learn More

Cell culture is at the heart of the production process for many biopharmaceuticals but finding the optimal conditions to maximize yield can be a complex and time-consuming process. Traditional process development relies on costly and labor-intensive set-ups, significantly limiting throughput and the range of experimental conditions that can be assessed.  Scientists in Roche Pharma Research and Early Development (pRED) have adopted an alternative approach, combining single-use microbioreactors with advanced automation and analytical platforms to streamline the workflow. Learn More

Many companies are working to advance the development of a vaccine in the context of the current COVID-19 crisis. New bioprocess technologies, such as single-use equipment and process automation, open up possibilities for quality control and validation. This is especially important in GMP-regulated environments, such as in the development and manufacturing of new influence vaccines. When facing a pandemic outbreak, the need to smoothly develop new processes and quickly scale up to clinical production volumes is key to efficiently develop new vaccinations. BioPharm International recently spoke with Dr. Jörge Schwinde, Key Segment Manager for Vaccines and Monoclonal Antibodies at the Bioprocess Center of Eppendorf, about the typical developmental process of new vaccines and how he COVID-19 pandemic influences the global vaccine developmental landscape. Learn More

Pseudomonas putida is a micro-organism with great potential for industry due to its stress-endurance traits and easy manipulation of the metabolism. However, optimization is still required to improve production yields. In the last years, manipulation of bacterial small non-coding RNAs (ncRNAs) has been recognized as an effective tool to improve the production of industrial compounds. So far, very few ncRNAs are annotated in P. putida beyond the generally conserved. In the present study, P. putida was cultivated in a two-compartment scale-down bioreactor that simulates large-scale industrial bioreactors. We performed RNA-Seq of samples collected at distinct locations and time-points to predict novel and potentially important ncRNAs for the adaptation of P. putida to bioreactor stress conditions.
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The emergence of cell and gene therapies (CGTs) as a novel therapeutic modality has ushered in significant scope and opportunity to address previously unmet need. The rationale behind such therapies is to use cells, somatic, or modified/reprogrammed, to trigger regeneration of damaged tissues or targeting of cancer cells, for example. With increasing numbers of CGT products receiving regulatory approval, there is now a focus on establishing scalable and robust manufacturing processes for both autologous (patient-specific) and allogeneic (universal donor) which are both cost effective and can meet expected demand. One of the cell types under evaluation in numerous clinical trials is human mesenchymal stem/stromal cells (hMSCs) that possess both immunomodulatory properties and in vitro differentiation capability. Learn More

It was developed a fed-batch bioprocess to produce a recombinant vaccine against Entamoeba histolytica under operational conditions attainable to large scale bioprocesses. We have produced this recombinant protein in shake flask and stirred tank bioreactor. Initial results in shake flask cultures under different methanol concentration of 0.5, 1.5 and 3% (v/v) produced extracellular protein at quantities of 10, 22 and 33 μg/mL, respectively. Then a scale-up process was performed from shake flask to fermenter by keeping similar volumetric power supply (P/V). The operational conditions were set up in fermenter as those used at commercial scale and supply of pure oxygen was avoided to keep the scalability of the bioprocess. Learn More

In this episode of How It's Made they explain how unlike traditional drugs synthesized from chemicals, biologic medicines are proteins made from living cells. Learn More

The success of genetically engineered T-cells modified with a chimeric antigen receptor as an adoptive cell immunotherapy and the subsequent last regulatory approvals of products based on this therapy are leading to a crescent number of both academic and pharmaceutical industry clinical trials testing new approaches of this "living drugs". The aim of this review is to outline the latest developments and regulatory considerations in this field, with a particular emphasis to differences and similarities between academic and industry approaches and the role they should play to coexist and move forward together. Learn More

There are many drivers to intensify the manufacturing of a biological product. Advances are occurring throughout the biomanufacturing arena, from process development techniques to improved manufacturing platforms, equipment and facilities. Many employ the term 'bioprocess intensification' to refer to systems for producing more product per cell, time, volume, footprint or cost. This need is being driven by two emerging priorities: cost control and process efficiency. We are seeing great interest in the power of such disciplines as synthetic biology, process simplification, continuous bioprocessing and digital techniques in the optimization of bioprocess development and manufacturing. Other powerful disciplines here include process automation, improved monitoring and prefabricated facility modularity and podularity. Learn More

In this future paper, we would like to take you on a journey into 2025. After all, we often don't realize what it is we should be doing until we look beyond what is right in front of us. Sluggish innovation and fear of disruption are of course common issues, however, allow us to set out in black and white exactly what we think needs to be done. Learn More

In this application note, we explain the differences between batch, fed-batch, and continuous fermentation and how these influence culture growth. As an example, we look at E. coli fermentation processes at bench scale. We track the biomass and nutrient concentrations during batch, fed-batch, and continuous fermentation runs. We explain different methods to analyze the process, including determination of biomass, growth rate, productivity, yield, and analysis of process costs. The comparisons can help bioprocess engineers to select the most appropriate method to meet their needs. In our examples we studied E. coli fermentation at bench scale. The principles may also apply to bioprocesses using other microbes or mammalian cells, at both smaller and larger scales. Learn More

Bioprocess scale-up is a critical step in process development. However, loss of production performance upon scaling-up, including reduced titer, yield, or productivity, has often been observed, hindering the commercialization of biotech innovations. Recent developments in scale-down studies assisted by computational fluid dynamics (CFD) and powerful stimulus-response metabolic models afford better process prediction and evaluation, enabling faster scale-up with minimal losses. In the future, an ideal bioprocess design would be guided by an in silico model that integrates cellular physiology (spatiotemporal multiscale cellular models) and fluid dynamics (CFD models). Learn More

Embryonic stem cells (ESCs) have almost unlimited proliferation capacity in vitro and can retain the ability to contribute to all cell lineages, making them an ideal platform material for cell-based therapies. ESCs are traditionally cultured in static flasks on a feeder layer of murine embryonic fibroblast cells. Although sufficient to generate cells for research purposes, this approach is impractical to achieve large quantities for clinical applications. In this study, we have developed protocols that address a variety of challenges that currently bottleneck clinical translation of ESCs expanded in stirred suspension bioreactors. We demonstrated that mouse ESCs (mESCs) cryopreserved in the absence of feeder cells could be thawed directly into stirred suspension bioreactors at extremely low inoculation densities. Learn More

The Covid-19 pandemic lead to a strong increase of vaccine development projects and new vaccine manufacturing approaches. In this panel discussion, experts from Eppendorf and ContiVir share their thoughts on how they are "Rethinking Vaccine Development". Learn More

The US FDA has released several guideline documents, which are likely to increase the adoption of digital technologies in pharmaceutical manufacturing. These guidelines provided frameworks for electronic submission of clinical data and manufacturing establishment information, while laying down expectations for maintaining data integrity and compliance with good manufacturing practices (GMP). These guidelines have encouraged pharmaceutical companies to adopt advanced processes such as modular manufacturing, electronic batch record systems and computerized maintenance management systems that employ sensors and advanced analytics. This serves as a great foundation for industry to shift from batch processing to continuous processing. Learn More

The convergence of DCS and PLC technologies has made it more challenging than ever for process manufacturers to select the best technology for their application. A successful evaluation should start with developing a clear picture of the requirements of your application and the needs of your engineering, maintenance, and operations personnel. In this paper we will explore key questions to help select the system that best meets your goals. We will demonstrate why having a clear picture of the application requirements and the needs of your engineering, maintenance, and operations personnel is paramount to finding the right automation technology. Finally, we will provide you with a checklist to help determine your optimal system. Learn More

The use of high-throughput systems in cell culture process optimization offers various opportunities in biopharma process development. Here we describe the potential for acceleration and enhancement of product quality optimization and de novo bioprocess design regarding monoclonal antibody N-glycosylation by using an iterative statistical Design of Experiments (DoE) strategy based on our automated microtiter plate-based system for suspension cell culture. In our example, the combination of an initial screening of trace metal building blocks with a comprehensive DoE-based screening of 13 different trace elemental ions at three concentration levels in one run revealed most effective levers for N-glycan processing and biomass formation. Learn More

How often do you pipette in your cell culture lab every day? Usually, we do it so often that we tend stop thinking about how to pipette right. With this educational webinar we would like you to stop for a moment, re-think what you are doing, and significantly improve your skills and results. We would like to share Eppendorf´s almost 60 years of experience in proper liquid handling with you - for more reproducible cell culture results and better contamination protection in your lab. Learn More

Being different from the continuous process, batch processes in the practical industry have several distinct characteristics, such as the unsteady state, severe nonlinearity, and iterative operation. For tracking a reference trajectory of a batch process, data-driven model predictive controllers have been proposed with the progress of sensors and machine learning. Among them, the latent variable space model-based controllers (LV-MPC) have been applied to the batch processes for decades. We propose to update the local model in the manner of just-in-time learning (JITL) and to use them to the predictive controller design at first. Learn More

Mammalian cell cultures are widely used in the biopharmaceutical industry to produce monoclonal antibodies, vaccines, growth factors, etc. Cell death is an essential biological process for physiological growth and development, but it is a major problem in biopharmaceutical production in bio-industry. Cell death within bioreactor occurs due to various intracellular and extracellular stresses. These stresses negatively affect the culture longevity, overall product quality, and yield. Among all cell death types, apoptosis accounts for most of the cellular death in the bioreactor. In this review, we classified and described different types of cell death and their molecular mechanisms and summarized the cell death inhibition approaches implemented to inhibit cell death for various applications. Learn More

Owing to the increasing demand for large scale and high efficiency in manufacturing processes, computer aided tools for process operation and control are rapidly gaining popularity. An important state variable in aerobic processes is the dissolved oxygen, which can be easily measured online and is an important indicator of the metabolic activity. However, due to the fast kinetics of the oxygen transfer, dynamical models describing aerobic bioprocesses tend to be highly stiff. In this work we use the slow-motion invariant manifold and the quasi steady state assumption methods to eliminate the differential equation describing the dissolved oxygen (the fast mode). Learn More

In recent years, multivariate data analysis (MVDA) and modeling approaches have found increasing applications for upstream bioprocess studies (e.g., monitoring, development, optimization, scale-up, etc.). Many of these studies look at variations in the concentrations of metabolites and cell-based measurements. However, these measures are subject to system inherent variations (e.g., changes in metabolic activity) but also intentional operational changes. It is proposed to perform MVDA and modeling on data representative of the underlying biological system operation, that is, the specific rates, which are per se independent of the scale, operational strategy (e.g., batch, fed-batch), and biomass content. Learn More

The adoption of next generation bioprocessing is challenged as biopharmaceutical companies grapple with the adoption of new manufacturing methods, which some consider to be essential to the industry's continued growth and innovation. Within the next five years, it is estimated that approximately 35 percent of today's biologics will be manufactured using some sort of process intensification methods. Facilities will be smaller but capable of producing higher volumes of multiple molecules.

The key to this advancement will be the use of methods such as continuous bioprocessing technologies, some of which may be single use, which would enable facilities to reduce both their size and cycle times. The complexity of the bioprocessing industry has challenged companies' innovation efforts, due to potential costs required for changes to take place. Learn More

In the biopharmaceutical industry, Raman spectroscopy is now a proven PAT tool that enables in-line simultaneous monitoring of several CPPs and CQAs in real-time. However, as Raman monitoring requires multivariate modeling, variabilities unknown by models can impact the monitoring prediction accuracy. With the widespread use of Raman PAT tools, it is necessary to fix instrumental variability impacts, encountered for instance during a device replacement. In this work, we investigated the impact of instrumental variability between probes inside a multi-channel analyzer and between two analyzers and explored solutions to correct them on model prediction errors in cell cultures. Learn More

A major hurdle during process development and optimization studies is the huge experimental effort in conventional design of experiments (DoE) methods. The combination of DoE with a virtual representation of the bioprocess, called digital twin, in model-assisted DoE (mDoE) can be used as an alternative to decrease the number of experiments significantly. In this chapter, digital twins and their role in mDoE are discussed. First, statistical DoE methods are introduced as the basis of mDoE. Second, the combination of a mathematical process model and DoE into mDoE is examined. This includes mathematical model structures and a selection scheme for the choice of DoE designs. Learn More

Human induced pluripotent stem cells (hiPSCs) hold enormous promise in accelerating breakthroughs in understanding human development, drug screening, disease modeling and cell and gene therapies. Their potential, however, has been bottlenecked in a mostly laboratory setting due to bioprocess challenges in the scale-up of large quantities of high-quality cells forclinical and manufacturing purposes. While several studies have investigated the production of hiPSCs in bioreactors, the use of conventional horizontal-impeller, paddle and rocking-wave mixing mechanisms have demonstrated unfavourable hydrodynamic environments for hiPSC growth and quality maintenance. This study focused on using computational fluid dynamics (CFD) modeling to aid in characterizing and optimizing the use of vertical-wheel bioreactors for hiPSC production. Learn More

Glutathione (GSH) plays a central role in the redox balance maintenance in mammalian cells. Previous studies of industrial Chinese hamster ovary cell lines have demonstrated a relationship between GSH metabolism and clone productivity. However, a thorough investigation is required to understand this relationship and potentially highlight new targets for cell engineering. In this study, we have modulated the GSH intracellular content of an industrial cell line under bioprocess conditions to further elucidate the role of the GSH synthesis pathway. Learn More

Biological molecules are widely produced by fermentation technology using bacteria, fungi or yeast. Fermentation is a biochemical process wherein the rate of bioconversion is governed by the organisms involved. The growth of the organism is mainly limited by mass transfer rates of nutrients and gases that directly affect the product formation in fermentation. The growth rate of microbes can be accelerated by increased mass transfer rates and cell wall permeability with the use of controlled low frequency ultrasound irradiation. The present review provides insights into the application of acoustic cavitation in process intensification of fermentation approaches and the role of various factors involved are highlighted with typical examples. Learn More

Ammonia is an ubiquitous compound. It is used by humans in industrial fertilizers and refrigerants. Furthermore, it is produced by animals as an excretion product as well as a product in amino acid degradation and waste decomposition. Continuous mammalian cell lines (CCLs), which are important hosts for the production of biological pharmaceuticals, generate ammonia during the cell metabolism. This is due to the lack of energy in form of ATP. To close this gap, these cells consume glutamine to produce α-ketoglutarate, which is further degraded in the mitochondria and as a second product poising ammonia. Ammonia inhibits cell growth and further yields to cell death. Learn More

The development of a continuous process for cell separation is growing rapidly due to the current trend of cost-effective manufacturing in biological industries. The continuous cell separation process has a significant reduction in capital equipment costs and facility size compared to the conventional batch process. In the study, a multi-layered microfluidic-based device integrated with the porous membranes was fabricated for continuous size-based isolation of the cells based on the mechanism of restrictive cross-flow filtration, allowing the biological sample entered in a single inlet of the device and separated into two outlet streams. Learn More

Due to its beneficial attributes, silicone is utilized today in countless medical and pharmaceutical applications. It is the preferred material for sealing complex systems as well as for peristaltic and pharmaceutical tubing. Silicone is available in a wide range of durometers, from super soft (000) to relatively hard (90 Shore A), and can withstand a range of temperatures. The silicones readily available in today's market differ not only in their level of approval for medical and pharmaceutical applications, but also in their crosslinking mechanism. There is a clear distinction between platinum-cured and peroxide-cured silicones. Learn More

In the past 15 years, the biopharma industry has seen a steady uptake in the use of single-use technology in bioprocessing. Single-use technology is being developed for nearly every application from upstream mixing and bioreaction to downstream perfusion and chromatography. In the pursuit of personalized medicine and continuous processing, single-use is being hailed as the only way to achieve these goals. Does this mean an end for traditional bioprocesses that employ stainless steel technology? Is stainless steel already being considered a legacy product as we journey towards fast, closed and process-intensified manufacturing systems?

Marc Pelletier, Director of CRB, and Mark Embury from ASEPCO, part of Watson-Marlow Fluid Technology Group (WMFTG), discuss whether stainless steel is a technology of the past. Learn More

This paper presents various software based approaches suitable for the design of knowledge-based monitoring of biotechnological production processes. These processes require special treatment with respect to the complexity in biochemical reactions which make the design and construction of reasonably complex and practically usable mathematical models rather difficult. Additional complexity arises from the lack of industrially viable sensors for on-line measurement of key process variables. Software sensors which often use tools from the field of artificial intelligence represent one of the suitable approaches for the overcoming of the above mentioned limitations for its ability to utilize effectively both quantitative and qualitative knowledge about the monitored bioprocess. Learn More

In the last years, regulatory agencies in biopharmaceutical industry have promoted the design and implementation of Process Analytical Technology (PAT), which aims to develop rapid and high-throughput strategies for real-time monitoring of bioprocesses key variables, in order to improve their quality control lines. In this context, spectroscopic techniques for data generation in combination with chemometrics represent alternative analytical methods for on-line critical process variables prediction. In this work, a novel multivariate calibration strategy for the at-line prediction of etanercept, a recombinant protein produced in a mammalian cells-based perfusion process, is presented. Learn More

What is between Downstream and Fill & Finish? Most of the time, fill and finish services for drug substances are not located at the same venue that the drug substance is produced at. Pharmaceutical and biotechnology companies look to contract manufacturing organizations (CMOs) as a way to meet their fill- and-finish needs and reduce risk. As a result, the shipping of liquid drug substances around the globe is often inevitable and the challenges arise. Learn More

The production of pharmaceuticals, industrial chemicals, and food ingredients from biotechnological processes is a vast and rapidly growing industry. While advances in synthetic biology and metabolic engineering have made it possible to produce thousands of new molecules from cells, few of these molecules have reached the market. The traditional methods of strain and bioprocess development that transform laboratory results to industrial processes are slow and use computers and networks only for data acquisition and storage. Digitalization, machine learning (ML), and artificial intelligence (AI) methods are transforming many fields - how can they be applied to bioprocessing to overcome current bottlenecks? Learn More

The various mammalian cell lines used for diagnostic and therapeutic protein expression exhibit distinct metabolic features. Even clones descending from the same parental cell line and expressing the same product can have individual nutritional requirements. We established a comprehensive platform for fast and robust quantitation of 55 cell culture media constituents and metabolites in spent media. This includes for example an advanced UPLC method developed in-house that allows for the parallel quantitation of more than 30 (non-) proteinogenic amino acids and dipeptides. Learn More

Based on conversations with customers over the past several years, Univercells has identified the major hurdles in largescale virus manufacture, which include expensive manufacturing facilities, lack of expertise, limitations in the number of scalable manufacturing technologies available in the market, and the high cost of good manufacturing practice (GMP) grade reagents including transfection mix, plasmids, and bovine serum. Incorporating design-of-experiment study principles and use of improved bioprocess control and data analysis software are allowing process development/optimization to be conducted at the multiparameter level... Learn More

Large-scale production of induced pluripotent stem cells (iPSCs) is essential for the treatment of a variety of clinical indications. However, culturing enough iPSCs for clinical applications is problematic due to their sensitive pluripotent state and dependence on a supporting matrix. In this review, we present an overview of cell culture media, suspension modalities, and monitoring techniques that preserve the quality and pluripotency of iPSCs during initiation, expansion, and manufacturing. Learn More

A continuous integrated bioprocess available from the earliest stages of process development allows for an easier, more efficient and faster development and characterization of an integrated process as well as production of small-scale drug candidates. The process presented in this article is a proof-of-concept of a continuous end-to-end monoclonal antibody production platform at a very small scale based on a 200ml alternating tangential flow filtration perfusion bioreactor, integrated with the purification process with a model-based design and control. Learn More

A survey of bioprocessing industry professionals asked which method they prefer for their connection needs: sealable and weldable tubing or aseptic connectors. Respondent opinion was almost equally divided with 52 percent choosing aseptic connectors and 48 percent preferring sterile tube welding. So how do you determine the right method to maintain batch integrity and maximize manufacturing flexibility while avoiding product loss, system downtime, and potential for operator error? Learn More

From MIT Video Productions comes "From Controversy to Cure," a 57-minute documentary history of the birth and boom of the molecular biology and biotechnology industries in Cambridge and specifically Kendall Square. Today, the area is home to one of the largest clusters of biotechnology and pharmaceutical companies in the world, and is, according to this film, "on the front lines to defeat COVID-19." Not long ago, Kendall Square was a wasteland of vacant plants and empty space cleared away for a NASA electronics research center project that was killed by the Nixon administration as part of spending cuts demanded by the crippling costs of the Vietnam War. Learn More

In biotechnology, the emergence of high-throughput technologies challenges the interpretation of large datasets. One way to identify meaningful outcomes impacting process and product attributes from large datasets is using systems biology tools such as metabolic models. However, these tools are still not fully exploited for this purpose in industrial context due to gaps in our knowledge and technical limitations. In this paper, key aspects restraining the routine implementation of these tools are highlighted in three research fields: monitoring, network science and hybrid modeling. Advances in these fields could expand the current state of systems biology applications in biopharmaceutical industry. Learn More

The purpose of this book is to present the different approaches most commonly employed in the control of bioprocesses. It aims to develop in some detail the bases and concepts of bioprocesses related to the control theory introduced in basic principles of mathematical modeling in bioprocesses. Learn More

Across the biopharmaceutical industry, the transition from traditional constructs such as large stainless-steel factories, batch manufacturing, and paper-based recording systems to modular facilities, single-use technologies, continuous manufacturing, and electronic systems is picking up pace. It's easy to see why: the benefits of greater agility and responsiveness for capacity planning, significantly reduced manufacturing costs, and improved product quality speak for themselves. Join Eliot Randle and Claire Hill as they discuss what's needed to address the current industry challenges and transform the future of biomanufacturing. Learn More

Process intensification strategies are needed in the field of therapeutic protein production for higher productivities, lower cost of goods and improved facility utilization. This work describes an intensification approach, which connects a tangential-flow- filtration (TFF) based pre-stage perfusion process with a concentrated fed-batch production culture inoculated with an ultra- high seeding density (uHSD). This strategy shifted biomass production towards the pre-stage, reaching up to 45 × 106 cells/ mL in perfusion mode. Subsequently, production in the intensified fed-batch started immediately and the product titer was almost doubled (1.9-fold) in an equivalent runtime. Learn More

This chapter introduces the necessary concepts to develop mammalian cell perfusion cultures for the expression of therapeutic proteins at lab scale. We highlight the operation of the orbitally shaken tubes and of a classical glass vessel reactor system coupled to an external alternating tangential flow (ATF) device. Two different experiments can be performed in the shake-tube system: (1) the VCDmax experiment exploring the maximum achievable viable cell density at a given medium exchange rate and (2) the VCDSS experiment for the prediction of process performance at constant viable cell density and a given medium exchange rate for the design of the benchtop bioreactor process. Learn More

We have developed an aptamer based microfluidic chip for inline monitoring of Ranibizumab in bioreactors. Aptamers for Ranibizumab were generated using 10 rounds of the SELEX process. Two of the aptamers exhibited high affinity towards the target analyte with a dissociation constant. The one with best affinity towards analyte was immobilized on gold micro-electrodes on a microfluidic chip, which was fabricated with glass base and PDMS top using conventional photolithographic technique. Immobilization steps were characterized using FTIR and EIS. Non-faradaic EIS measurement was used for label-free detection of Ranibizumab. Learn More

This report summarizes linearity, precision, and comparability of the new generation BioProfile FLEX2 to the previous generation BioProfile FLEX cell culture analyzer. As part of the internal validation for BioProfile FLEX2, sample replicates for all measurable parameters (pH, pCO2, pO2, glucose, lactate, glutamine, glutamate, ammonium, sodium, potassium, and calcium) were analyzed across the full analytical range on both BioProfile FLEX2 and BioProfile FLEX. Twenty replicate samples of control material were also analyzed consecutively on BioProfile FLEX2 to determine precision. BioProfile FLEX2 provided a high degree of precision, accuracy, and comparability against BioProfile FLEX analyzer. BioProfile FLEX2 exceeded all validation criteria for release. Learn More

In pharmaceutical production, only sensors that meet the highest hygienic standards are suitable for measuring the flow of products, from cleaning solutions to barely conductive water for injection. According to a study, Coriolis and SAW sensors both ensure high quality flow rate measurements but differ in terms of the level of maintenance required and the price/performance ratio. Learn More

In cell line development the identification of stable Chinese hamster ovary cells for production is a critical but onerous task. The stability trial focus upon high-level attributes can mask profound underlying cellular changes, leading to unstable clones mistakenly being chosen for production. The challenge is to assay underlying cell pathways and subsystems without pushing up cell line development costs. ChemStress® cell function profiling is a simple, multiwell plate-based assay that uses a panel of active chemicals to mimic known bioprocess stresses and challenge key pathways... Learn More

Cell density and viability are critical process parameters in many cell culture applications for biologics development and production. Monitoring the health and growth of the culture is so important that numerous methods have been developed to meet this need. Automated cell counting has become the gold standard in many applications due to its consistency, accuracy, and ease of use. Additionally, by trypan blue dye exclusion method some systems integrate directly with bioreactors or microbioreactor systems to provide on-line measurement, streamlining workflow. Learn More

Multiple process analytical technology (PAT) tools are now being applied in tandem for cell culture. Research presented used two in-line probes, capacitance for a dynamic feeding strategy and Raman spectroscopy for real-time monitoring. Data collected from eight batches at the 15,000 L scale were used to develop process models. Raman spectroscopic data were modelled using Partial Least Squares (PLS) by two methods-(1) use of the full dataset and (2) split the dataset based on the capacitance feeding strategy. Root mean square error of prediction (RMSEP) for the first model method of capacitance was 1.54 pf/cm and the second modelling method was 1.40 pf/cm... Learn More

A novel functional micropipette tip for DNA purification, called DNAtip, is described based on a novel coating process. The functional layer is prepared by an internal coating with silica particles supported into a polyvinyl alcohol (PVA) film. The coating layer is stabilized by simultaneous radiation-induced crosslinking to avoid its dissolution. Three types of silica particles of different diameters: micron, sub-micron, and nano sizes are studied. In addition, pH of the coating solution and concentration of the silica particles were optimized. Tips coated with fumed silica showed the highest absorption capacity of a plasmid DNA with an acceptable purity. Learn More

Process intensification in mammalian cell culture-based recombinant protein production has been achieved by high cell density perfusion in recent years. As the majority of therapeutic proteins are produced in Chinese Hamster Ovary (CHO) cells, intensified perfusion processes have been mainly developed for this type of host cell line. However, the use of CHO cells can result in non-human posttranslational modifications of the protein of interest, which may be disadvantageous compared with human cell lines. In this study, we developed a high cell density perfusion process of Human Embryonic Kidney (HEK293) cells producing recombinant human Erythropoietin (rhEPO). Learn More

In this work, anchorage-dependent baby hamster kidney cells (BHK-21) were cultivated on microcarriers. Growth was monitored with off-line cell counts compared with on-line viable cell density monitoring based on permittivity. The two methods correlate with an R2of 0.99 in exponential phase. Therefore, permittivity-based viable cell density monitoring can be employed to reduce the effort and risks of off-line sampling. Learn More

In this work, a Vero cell line used to produce viral vaccines was used by Intravacc to perform the cell and virus cultivations in Applikon's newly developed small-scale customizable single-use bioreactors. The growth curves of Vero cells, were compared with the growth curves of Vero cells growing in conventional autoclavable glass bioreactors under the same conditions and in the same culture volume. Subsequently, a virus for which vaccines are needed, EV71_C4, was grown on Vero cells. The single-use bioreactors are suitable for Vero cell culture and EV71_C4 virus propagation because there was no difference with respect to Vero cell culture and EV71_C4 virus culture between the glass bioreactor and disposable bioreactor. Learn More

A long-standing challenge for on-line optical cell biomass probes has been to maintain biomass prediction accuracy across changing process conditions, particularly for organisms requiring highly variable rates of agitation and aeration in order to avoid oxygen-limited growth. For microbial organisms grown to high cell biomass densities, being able to accurately track the biomass across the entire growth cycle from inoculation to harvest has been difficult to achieve due to the limited linearity range of traditional on-line methods and variation in sensor readings with changing process conditions. A comparison study of the linear range and the effects of changing agitation and aeration conditions on biomass prediction error for a methanotrophic culture are compared for two commercially available on-line optical reflectance probes. Learn More

This 20-page white paper authored by Hamilton provides a comprehensive, but accessible, overview of requirements and associated recommendations related to the FDA's Process Analytical Technology (PAT) initiative for the biopharmaceutical industry. Learn More

Technology transfer is a key milestone in the journey from discovery to full-scale GMP-compliant manufacturing. Pall's combined knowledge and understanding of the processes, and technological capabilities will help you navigate through the challenges of your tech transfer reducing your costs, resources and time. Download the BPI article and learn how partnering with Pall can help you navigate through the challenges, to achieve a successful tech transfer. Learn More

Shaken cultures of suspension-adapted animal cell lines are still the method of choice for screening and small-scale culture experiments due to their easy handling, the simple and fast implementation in almost any laboratory environment and the possibility of cost-effective parallelization. However, on closer inspection, there are a few points that need to be considered and which play a major role in successful cultivation. Learn More

Protein freeze-thawing is frequently used to stabilize and store recombinantly produced proteins after different unit operations in upstream and downstream processing. However, freeze-thawing is often accompanied by product damage and, hence, loss of product. Different effects are responsible, including cold denaturation, aggregation effects, which are caused by inhomogeneities in protein concentration, as well as pH and buffer ingredients, especially during the freeze cycle. In this study, we tested a commercially available small-scale protein freezing unit using immunoglobin G (IgG) as monoclonal antibody in a typical formulation buffer containing sodium phosphate, sodium chloride, and Tween 80... Learn More

Critical process parameters (CPPs) are the key variables in the pharmaceutical manufacturing process that are most likely to affect the quality attributes of a biological product. CPPs must be monitored and controlled throughout the various stages of production. The most common CPPs measured are pH, dissolved oxygen, conductivity, capacitance, and biomass. In this white paper from CPC, the authors discuss the role of traditional sensors in single-use systems including the benefits they offer, their associated challenges and the solutions available for their successful use. Learn More

Biomanufacturing relies on numerous pieces of equipment working in concert to produce life-altering therapeutics. The equipment relies on various subsystems to achieve the desired results. In a bioreactor, one of the most essential subsystems provides gas management for the gases necessary for cellular metabolism. At the heart of the gas management subsystem is the thermal mass flow controller (MFC), a component that precisely measures and controls the delivery of gases to the bioprocess. Learn More

The development of bioreactor processes for cGMP monoclonal antibody (mAb) manufacturing requires reliable, accurate, and timely in-process analytical data to guide process development decisions. The measurement of IgG expression level or titer is a key upstream process output that can present challenges for the rapid and systematic optimization of robust, scalable process. This study compares Protein A HPLC with the Roche Cedex Bio instrument for profiling the expression of multiple antibody products generated from CHO cell bioreactor culture. Considerations of comparative instrument performance, accuracy, economics, and versatility are presented in the context of accelerating development timelines from bench scale development to manufacturing clinical antibody supply. Learn More

In this application note, published by ForteBio, the analytical group at Boehringer Ingelheim, Fremont, USA illustrates how they needed a robust assay to measure the biological activity of an antibody fragment (Fab) molecule for in-process testing as well as stability and lot release testing in their Quality Control (QC) department. The group was able to develop a working Fab activity assay on the Octet® RED system in less than a week. Learn More

The economically competitive bioproduction of specialty chemicals using microbial cultures often demands the use of complex feedstocks that have undergone minimal processing and purification. Such feedstocks may be high in cellulosic content, have dark color, contain a wide mixture of particle sizes, and exhibit substantial batch-to-batch variation. The real-time monitoring of cell density (or cell biomass) is essential to maintaining the health of the microbial culture and for optimizing the yield of bioproducts, but has been a long-standing challenge within such complex feedstocks. Learn More

The economically competitive bioproduction of specialty chemicals using microbial cultures often demands the use of complex feedstocks that have undergone minimal processing and purification. Such feedstocks may be high in cellulosic content, have dark color, contain a wide mixture of particle sizes, and exhibit substantial batch-to-batch variation. The real-time monitoring of cell density (or cell biomass) is essential to maintaining the health of the microbial culture and for optimizing the yield of bioproducts, but has been a long-standing challenge within such complex feedstocks. Learn More

In this third installment, Dr Aditya Bhat and Dr Sven Ansorge discuss the usefulness of capacitance measurement in Viral Vector-vaccine production processes, highlighting Sven's work and pieces of research for this pertinent topic considering the current Covid-19 pandemic and also looking at the adoption of capacitance in cGMP manufacturing with reference to the useful papers produced by Biogen in recent years. Learn More

ABEC is inviting you on a live guided tour through their CSR® Single-Use Process area, CSR Disposable Container (DC) Manufacturing ISO-7 clean rooms, and the bioprocess equipment manufacturing floors of their USA & Ireland Facilities. Learn More

On-line parameters like pH and dissolved oxygen are commonly used to optimize process yield. Further optimization is now possible with a new breed of sensors for on-line measurement of viable and total cell density. Data from traditional off-line measurements is limited and time consuming to obtain. The off-line results are delayed, and process adjustments are manual. Real-time measurement and control based on cell density is a critical consideration for any modern automated control strategy. Learn More

This video, produced by Applikon, illustrates their offering of tailor-made 500mL and 3L single-use bioreactors. By using 3D printing technology they are able to create any bioreactor configuration that is required for your optimal cell culture or fermentation process. Learn More

In Part 2 of this 3 Part series, Dr Aditya Bhat and Dr Sven Ansorge take a closer look at the speed of adoption of capacitance for measuring biomass in-line compared to other parameters and the impact of the FDA's PAT initiative on accelerating the adoption of capacitance as a really useful tool in manufacturing. Learn More

In this project, oxygen mass transfer in liquids through the use of different bubble generation equipment is examined. The purpose of this project is to first investigate the bubble formation through different type of spargers made from different materials and different governing oxygen transfer phenomena. Based on the analysis of the investigations, equipment weaknesses that might be crucial for some applications are identified and finally, an equipment design, which eliminates the identified weaknesses, is proposed and validated. Learn More

This new eBook, published by industry leader Brooks Instrument, takes you through the key steps to ensure your approach to calibrating bioreactor MFCs is accurate and on target. Understanding the role calibration plays in long-term MFC performance - and what steps you should take to ensure proper calibration - can help sustain bioprocessing results. Learn More

In the first installment of this three part interview series produced by Aber Instruments, Dr Aditya Bhat interviews Dr Sven Ansorge, exploring Sven's background, his experience and Sven's insights on the development of capacitance technology over the years, looking at it's adoption within the bioprocess industry and how capacitance has evolved to become a useful tool for monitoring biomass and controlling critical parameters, relating to the PAT initiative from the FDA. Learn More

The mass transfer coefficient (kL)and the value of the interfacial area (a) is used to characterize bioreactor systems for their oxygen transfer capability. The kLa value and the mixing time are amongst the most important performance indicators for bioreactors. Their calculation and modeling is extremely complex in highly aerated systems. This paper describes a new methodology for measuring the kLa value to accurately determine the coefficient. The valuable insights obtained allow for improved bioreactor characterization, thus, making it easier to meet regulatory requirements. Furthermore, examples outlining the benefits in the practical application of the new methodology are presented. Learn More

This case study, authored by AdvantaPure, illustrates the performance of their custom BioClosure systems for bulk drug substance. The primary objective of the customer was to develop a closed system that was leak proof under operating conditions and maintained container closure integrity during storage at -70oC. Additionally, the customer was concerned about foaming that occurred when filling the container. AdvantaPure's engineering team worked with the customer to design a custom system for their production needs. Learn More

What would it take to rapidly manufacture 3 metric tonnes and then 10 metric tonnes of a mAb? Optimized product development procedures to speed up the transition of a discovered mAb into clinic and into commercialization have been discussed in an excellent article by Brian Kelley. What about subsequent commercial manufacturing capacity? According to Ecker and Seymour, 23 metric tons of approved biologics were produced in 2018, with 65% of that capacity controlled by 10 companies. Squeezing an extra 10% of the global capacity out of the network seems achievable, particularly if these top 10 companies are involved. We used Biosolve Process to estimate the current manufacturing capacity and timeframes for production of these products. Learn More

Continuous processing is already well accepted on the upstream processing side, since perfusion mode culture has been in use for decades at an industrial scale.Justifications for switching from batch to continuous chromatography are numerous. Besides, the rising demand of biologics suggests reduced processing and labor costs, footprint reduction, flexibility increase, more stringent requirements for controlled, more consistent and improved quality; and most importantly, the requirement for higher productivity and therefore overall manufacturing cost reduction. This white paper, authored by Novasep, highlights why and how to intensify your bioprocess. Learn More

This video, produced by AdvantaPure, shows the burst, leak/integrity and vacuum testing of their AdvantaSil™ High Pressure Silicone Tubing by the development team. Learn More

This white paper, authored by Nordson Medical, explains the role these new manifolds play in simplifying processing applications by offering companies an option that reduces the number of connections, shut-offs, and disconnections needed in typical production processes. The new component replaces older assemblies like ganged stopcocks and multiple leg manifold arrangements of fittings and pinch clamps. Learn More

Purification of enveloped viruses and virus-like particles presents several challenges due to their large size and complexity. Here we present a case study about resin screening, process development, and scale-up purification of a retrovirus-like particle. The resulting cGMP- compatible process required approximately 4 hours to purify 240 L of nuclease-treated bioreactor harvest and resulted in a 99% reduction in process volume and up to 65% recovery of virus particles with a final purity consistent with requirements for clinical trials. This protocol is readily scalable, fast, low cost, simple, and cGMP-compatible and can be used for the capture step in clinical-grade manufacture of retrovirus vectors. Learn More

This handbook, created by Cytiva, describes the use of size exclusion chromatography (SEC) for the purification and separation of biomolecules, with focus on practical information for optimized results. Since the introduction of the first SEC resin, Sephadex in 1959, SEC has played a key role in the purification of proteins and enzymes, polysaccharides, nucleic acids, and other biological macromolecules. Learn More

In this well written article from PharmTech, the author explores how modular manufacturing is being increasingly used in the biopharma industry where flexibility and low-cost manufacturing systems are sought out. The design of a modular system in biopharma processing can involve breaking down a manufacturing facility into smaller functional building blocks, also known as modules. Learn More