Don’t miss our top articles of 2020! I have compiled a list of our 25 most popular articles for 2020 in alphabetical order.

3D Model Systems: Spheroids, Organoid and Tissue Model Systems

Culturing cells outside of their natural environment in a laboratory under controlled conditions has become essential to scientific research. Cell culture has utility in diverse areas from stem cell and cancer research, monoclonal antibody production, drug discovery, regenerative medicine, therapeutic protein production and for modeling diseases. Cells to establish in vitro cultures can be isolated from normal or diseased tissues, be grown as adherent monolayers or in suspension, and can be established in two or three dimensions…

Baculovirus Expression System Demonstrates High Yield Protein and Viral Vector Production from Micro to Large Scale

Vaccines and gene therapies have a critical role to play in the future of medicine. As seen with the current Covid-19 pandemic, rapid development and manufacture of vaccines and viral vectors can have a tremendous impact on our ability to address and treat emerging healthcare needs. Scalable and efficient protein and viral vector production are essential to the development of vaccines and gene therapies. However, this often presents challenges, as expression systems don’t provide the versatility required to meet multiple formats or production requirements…

Best Practices for Rapid High Titer Protein Expression

Recombinant protein expression is critical to research, biotherapeutic drug discovery and structural studies. Choosing the right protein expression system is key and the specific requirements of each application should be carefully considered. For instance, protein solubility, functionality, time to material, and required yield are often important considerations when selecting expression method…

Best Practices for Successful Technology Transfer

To achieve a successful technology transfer, both parties must commit to creating a detailed plan that will anticipate potential issues and provide a clear direction for exchange of information and ongoing communication. While there is always a desire to move as quickly as possible, the time and effort spent mapping out the logistics for technology transfer can save significant time and resources in the long run. Planning should include all process areas including identification of raw materials, equipment sourcing, resource availability and ongoing delivery schedules. There should also be a designated team with members from both the partner and client with routine communication and a planned training schedule…

Bioprocess Intensification – A discussion on the what, the how, and the why

Bioprocess intensification is a term that we hear often, but it is difficult to define or rather there are often several definitions depending on the perspective of the author. That said, it is generally considered a toolbox of strategies to improve the biomanufacturing process and make it more efficient. What is meant by “efficient” is a metric that has to be defined by the group working to intensify the process. It could be increasing productivity, lowering manufacturing costs, better utilization of facility space, reducing timelines, improving product quality or all of the above and more…

Design of Experiment Principles to Optimize Benzonase® Endonuclease in Viral Gene Therapy Applications

For over 30 years, Benzonase^® endonuclease has been used for the removal of nucleic acids in virus vector and vaccine manufacturing as mandated by regulators to ensure the purity and safety of the final product. The enzyme can degrade both DNA and RNA into small 3–5 base pairs fragments (<6 kDa) with no base preference, making it a versatile tool for process development to optimize the yield in virus purification as well as to improve the efficiency of downstream chromatography and filter devices. However, Benzonase^® endonuclease activity is strongly influenced by several factors (Figure 1) and balancing these is a critical step during process development to maximize its benefit in the overall workflow…

Flexible Platform Solutions Overcome Vaccine Production Challenges

This article explores the challenges of vaccine development and how they can be overcome by applying a quality by design (QbD) platform approach from early process development phases forward. The impact of the COVID-19 pandemic on vaccine production is evaluated, with insight on how flexible, integrated, closed-system processing is advancing the future of safe and efficacious vaccine production…

How a Next Generation Proteomics Platform is Accelerating the Fight Against COVID-19

Since the beginning of the COVID-19 pandemic, researchers from around the world have been working tirelessly to understand the biological pathways of the disease and develop effective therapeutics. One important enabling approach is proteomics. Proteomics is broadly defined as the study of proteins. A proteome is an entire set of proteins produced in an organism during its life cycle or at a given moment in time. In medicine, proteomics is used to provide insight into health and disease and increase our understanding of biology in real-time…

Important Considerations for Reducing Risk of Viral Contamination in Biologics, Cell and Gene Therapy Manufacturing

A study conducted through a consortium of biotech companies and the Massachusetts Institute of Technology’s Center for Biomedical Innovation provides valuable evaluation of past contamination events and best practices for reducing risk. Biologics manufacturing has a long history of safety. However, the use of cell culture to produce recombinant proteins does carry some risk of contamination either through bacterial or viral contaminants. While these events have been rare, they come at a very steep price when they do happen. The cost of a contamination can easily reach tens of millions of dollars and can cause drug shortages that negatively impact patients. For the biopharmaceutical company, it can also mean the loss of competitive advantage; a reduction in valuation and extensive remediation with facility shut downs. Thus reducing contamination risk is a high priority and requires regular study to improve prevention and detection methods…

Insights on Successful Gene Therapy Manufacturing and Commercialization

The first approved gene therapy study was conducted by the National Institutes of Health (NIH) in 1989 and provided evidence for the first time that human cells could be genetically modified and returned to the patient without harm. Since then, the industry has grown significantly with 22 gene and gene-modified cell therapies approved by regulatory bodies from various countries as of August 2019. As of October 2020, six gene and gene-modified cell therapies have been approved for use in the United States, including the latest approval in July 2020 for the gene therapy Tecartus™…

Intensified and perfusion microfluidic bioreactor with stirred tank performance in just 2 mL working volume

Scaled down models are becoming increasingly important to cell line and media development, as well as overall process optimization. These models provide the opportunity to run multiple experiments in parallel at a fraction of the time and cost it would take to run larger scale experiments. In many cases, without scale down models, incredibly informative multi-factorial experiments would not be feasible…

Media Optimization Strategies for Modifying Monoclonal Antibody Quality Attributes

Cell culture media optimization continues to play a critical role in both improving titers and providing a mechanism for modifying monoclonal antibody (mAb) quality attributes. Furthermore supplements and trace contaminants in the media can have dramatic effects, both positive and negative, on productivity, growth and protein quality. This highlights the importance of identifying media components that drive both productivity and desired quality profiles to select the best media formulation for each monoclonal antibody product…

Modernize Your Cell Development Assays with High-Throughput Image Cytometry

Cell line development encompasses an optimized process that balances cell line productivity and cell product quality over a consistent and reproducible timeline. This article summarizes areas of cell line development where image cytometry has provided workflow advantages and made the entire process more efficient. There are several solutions available for research groups within cell line development. For most manufacturers, CROs, and larger scale operations, there can be a daily need to process stacks of plates utilizing a high-throughput solution. As bioprocessing moves towards high-throughput and automation solutions – it seems prudent to replicate already established methodology from leaders in cell line generation…

Optimizing Characterization and Quantification of Therapeutic Oligonucleotides

Oligonucleotides (OGN) are a class of therapeutics comprised of short strings of synthetic nucleotides. OGNs have the ability to to alter, suppress or restore expression of target genes associated with disease or health. This makes them uniquely positioned to provide treatment for diseases with a known genetic component. There are currently more than 80 companies currently investigating OGNs to treat diseases and it is a therapeutic modality that is growing dramatically. The first OGN therapy, Vitravene also known as Fomivirsen, was approved in 1998. Since then, the application of OGNs has continued to evolve and several additional approvals have followed. As is the case with most emerging therapies, there are manufacturing challenges, in particular characterizing and quantifying OGNs can be difficult…

Organoid Models: Getting Started and Culture Tips

An organoid is defined as an in vitro 3D cellular cluster derived from primary tissue (lineage restricted adult stem cells), embryonic stem cells (ESCs), or induced pluripotent stem cells (iPSCs), capable of self-renewal, self-organization, showing similar organ functionality as the tissue of origin. Their similarity in composition and function to or-gans in vivo makes them invaluable 3D models to study organogenesis, disease modeling, and patient-specific therapies. Organoids recapitulate a large number of biological parameters including the spatial organization of heterogeneous tissue-specific cells, cell-cell interactions, cell-matrix interactions, and certain in vivo physiological functions generated by tissue-specific cells within the organoid¹. Organoids bridge a gap between 2D and animal model systems by providing a stable system amenable to extended cultivation and manipulation. As such, researchers have created physiologically relevant organoid models for many organs. That said, working with organoids is not without its challenges. High quality cell culture tools are important to ensuring success when researchers are looking to incorporate organoids into cell culture workflows…

Platform Approach to Accelerating Cell Line and Process Development

Cell line development and process development are critical building blocks for successful biomanufacturing. For cell line development, high performing clones must be identified as quickly and efficiently as possible. However, the reality is that traditionally this process has been labor intensive and time consuming. The clone selection process involves looking for a small number of clones that are expressing sufficient titer of the recombinant protein to become a candidate for production scale. To find these high performing clones often requires screening of thousands of candidates, either manually or through automation. After the high quality clones are selected, process development begins. Process optimization for these clones and specific application is conducted, which can also be time consuming and labor intensive…

Practical sustainable processing of post-use plastics in an increasingly single-use biomanufacturing environment

The benefits of single-use technologies in biomanufacturing are undeniable. Single-use products minimize operational risk, personnel requirements and validation activities. They also reduce utilities and water cost in cleaning and sterilization, as well as batch changeover time. This last factor is particularly valuable in pilot and small-scale manufacturing where frequent changeover is often necessary. And, as more personalized products become approved, we will no doubt be seeing even smaller manufacturing lots become an increasing priority…

Qualification of a Rapid In-house PCR-based Mycoplasma Assay for Screening of Cell Therapy Products

Cell therapy products require microbial testing including, mycoplasma, sterility, and endotoxin analysis as part of the final product release criteria for patient safety starting in Phase I. This is a regulatory requirement as product contamination can create real risk for patients, particularly if patients are pediatrics, geriatrics or immunocompromised. However, cell therapy products have unique properties, which make using traditional compendial microbial methods difficult or even impractical. This is especially true for mycoplasma testing where the conventional compendial culture (broth and agar) method requires a large testing volume, takes 28 days for results, and is not able to be easily automated or scaled. These are especially challenging in cell therapy products, where products do not have a long shelf life and production volume is often limited. In addition, in the case of CAR T products, patients are often being treated with these therapeutics as a last option and as a result, timing is crucial. Thus a short manufacturing length, including quick quality control testing turnaround times for these therapeutics can be critical…

Rapid Mycoplasma Testing for Biologics – FAQs

Cell and gene therapies are poised to revolutionize healthcare, however manufacturing of these products poses new challenges for quality and safety assurance strategies. This is especially true in testing for mycoplasmas, an ubiquitous and practically invisible bacterial contamination of cell cultures. These challenges were a common thread in presentations at the 2019 PharmaLab Pre-Conference Workshop on Mycoplasma qPCR Testing, especially given the anticipated revision of European Pharmacopoeia chapter 2.6.7. Consensus points included, the importance of using cellular matrices for testing and the potential of an implementation strategy that positions NAT as a fast first line of detection, followed by results verification via culture-based compendial methods. In an effort to investigate some of the most frequent challenges and provide useful answers to frequently asked questions, we spoke with experts and highlighted several consensus points from conference presenters and audience members…

Rapid Spent Media Analysis of Metabolites During Cell Line Development for Better Product Quality Outcomes

Spent media analysis is an important tool used to provide crucial information about the amino acids and nutrients that cells use during growth phase. Spent media analysis also provides details about the metabolites that are produced as a result of cell growth. This is useful to cell line and process development teams in that it provides data on media composition performance, optimal harvest time and product quality changes that can occur during culture…

Scalable High Yield System for Mass Culture of Attachment-Dependent Cells

As increasing numbers of cell and gene therapies progress from research to clinical trials and on to commercialization, the need for efficient manufacture of high numbers of cells is critical. Many of the cells used in these therapies are attachment-dependent, which means that they need a surface on which to grow and expand. This need for attachment makes growing them in large numbers more challenging, as adherent systems do not traditionally scale up to the high volumes that can be achieved with suspension systems. However, suspension systems typically require more time and resources to develop a successful process…

Single-Step Cloning Strategy Speeds Timeline for Novel Vaccine Candidates

With the emergence of Covid-19 this year and the threat of other novel viruses looming on the horizon, now more than ever we need fast, efficient methods for generating novel vaccine candidates. These new methods could be developed as new technologies, rethinking existing technologies, or combining existing technologies in a new way. I am pleased to share the following guest blog, which presents a method developed by GSK for single-step cloning to accelerate the novel vaccine development timeline. The approach combines existing technologies to shorten the cloning process for CHO cells to a single-step…

Spheroid Models: Getting Started and Culture Tips

It has been well established that three-dimensional (3D) cell culture is better at replicating the physiological interactions between cells and their environment than traditional two-dimensional (2D) cell culture. In fact, many types of mammalian cells have the ability to self-aggregate into 3D aggregates called cellular spheroids when cultured in suspension or in a nonadherent environment. Common examples of spheroids include embryoid bodies, mammospheres, hepatospheres, and neurospheres. Perhaps the most widely utilized spheroid model is the multicellular spheroid model (MCTS) formed from cancer cells, which is commonly used as avascular tumor models to gain mechanistic insight into cancer invasion and metastasis and as an anti-cancer therapeutic screening tool…

Successful Exosome production using Stem Cell Culture in Stirred Tank Bioreactors

Exosomes have been increasing in popularity as a possible therapeutic. They are being explored as an alternative to stem cell therapy for tissue and organ restoration. Exosomes are naturally occurring, mobile, membrane-limited, 30 – 100 nm in diameter, extracellular vesicles containing a large number of proteins, lipids, messenger, and micro-RNAs. They have been shown to play a role in the mediation of intercellular communication, the modulation of immune-regulatory processes, tumor metabolism, and regenerative as well as degenerative processes…

Successful Techniques for Progressing Biotherapeutic Candidates from Discovery to the Clinic

The reality is that many initially promising biotherapeutic candidates never make it to commercialization. It is estimated that only 1 in 1000 preclinical candidates reach the commercial market. Why does this happen? There are many that fail due to lack of efficacy or safety, but there are also candidates that fail due to stability, aggregation, and other issues related to cell line or process issues. While some issues may never be satisfactorily resolved to the point where commercialization is possible, others can be saved with early identification, engineering and process development approaches. Even for candidates that can’t be rescued, it is critical to know this early, so that time and resources aren’t wasted on a product that ultimately has no path forward. Therefore, it is imperative that strategies and technologies are available to evaluate and identify issues early in the development phase and address challenges where possible…

Viral Vector Process Intensification: Stepwise Model for Efficient Scale-up

The unprecedented clinical success of novel cell and gene therapies (CGT), such as CAR-T cell therapy, has led researchers to shift from treating more rare conditions present in smaller populations to those more prevalent in the general population. This has, in turn, resulted in a growing market demand for the viral vectors used in these therapies. It has highlighted the need to increase manufacturing capacity to meet market demands by developing scalable and cost-effective viral vector manufacturing processes and improving methods for purification and analysis to ensure product purity, efficacy, and safety. Research and Markets recently reported the global viral vector manufacturing market was valued at US$459.4 million in 2019 and – expected to reach US$2.2 billion by 2027…