Don’t miss our top articles of 2021! I have compiled a list of our most popular for 2021 in alphabetical order.

3D Bioreactor Platform Provides Increased Process Control and Throughput for Spheroid and Organoid Generation and Culture

Conventional adherent two-dimensional (2D) tissue culture where cells are grown on planar, rigid substrates has been the basis for many scientific discoveries. However, as cell culture techniques have advanced to grow cells in three-dimensional (3D) space, comparative studies with 2D cultures demonstrate they are simply unable to recapitulate the complex tissue architecture and cell connectivity observed in vivo. This can limit their utility to model cellular morphology, viability, proliferation, differentiation, gene and protein expression, and responses to drug and toxic compounds. Because of this, biomimetic 3D cell culture models, such as spheroid and organoids have gained broader acceptance for translational research, drug discovery and toxicology programs…

The Art and Science of Cell Food: How smart media formulations drive performance

The old adage ‘you are what you eat’ could equally be said for cell culture, because the quality of what you put into your bioreactor dictates the quality of what comes out. Several major challenges with cell culture media and processes must be overcome to unlock the full potential for successful discovery and efficient production of biopharmaceuticals. Many of these challenges are now being addressed through a unique and innovative portfolio of dipeptides that boost performance. Cell culture processes are the technological foundation of the biopharmaceutical industry. The continued revolution in health care, driven by biological drugs like antibodies and vaccines, has raised the importance of high-performing cell culture for the biopharmaceutical industry…

Best Practices – Cell Culture Techniques

The ability to take cells from their natural environment and culture them artificially in the lab has been an invaluable tool for researchers to study cell physiology, metabolic pathways as well as look at cellular responses to drugs and toxic compounds. Additionally, cells cultured in vitro have been successfully utilized for the large-scale production of biological compounds (e.g., vaccines, therapeutic proteins). That said, culturing cells in the laboratory is not a trivial task and success is dependent on a number of factors ranging from having the proper equipment, sterile technique, culture media and conditions, to utilizing the appropriate protocols for handling the cells during passaging and freezing/thawing…

Cell Culture Advancements and Opportunities for Future Improvement

Culturing cells in the lab for the purpose of research and biologics production is no easy task. It requires success across a number of areas including optimized culture media and conditions, proper equipment, good technique and efficient protocols. Cell culture media optimization plays a crucial role in increasing productivity, ensuring cell growth and health, and even as a lever to affect critical quality attributes. Thus, cell culture scientists have an essential role in advancing the discovery and production of existing and new therapeutic modalities…

Cell Culture Media Analysis for Cell Therapy Applications

In the field of regenerative medicine, cell therapies are increasingly becoming viable therapeutic alternatives to traditional treatments for a wide range of diseases. Most recently, T-cells have been the focus of much clinical investment with chimeric antigen receptor (CAR) T-cell immunotherapies for blood cancers, but other cell types including mesenchymal stem cells (MSCs) and other immune cells (i.e., NK cells, macrophages) have also been investigated for their therapeutic potential. Regardless of the cell type, understanding the best way to culture the cells in vitro to achieve optimal cell activation and expansion, while maintaining functionality is key to the success of these “living medicines”…

Cell Culture Media Analysis for Gene Therapy

The genes within our cells contain the necessary information to direct the production of proteins and enzymes to support normal functions in the body. However, when a gene or part of a gene is defective, mutated or missing, this can disrupt normal functions manifesting in a number of health problems and diseases. Gene therapy aims to treat diseases through genetic modification, most frequently genetically engineered viral vectors are used to deliver a genetic payload to the cells. There are non-viral methods in development, however, viral vectors are still the most popular approach with two-thirds of the clinical trials to date delivered via viral vector.¹ It has the potential to transform medicine and create new therapeutic options for patients who are living with difficult, and even incurable, diseases. The gene therapy can be delivered directly in vivo to affected cells through either providing the cell with working copies of the gene or by silencing mutated genes that function improperly and cause disease. Another gene therapy strategy aims to reengineer cells ex vivo to increase their therapeutic efficacy against a target population. This is the principle behind CAR T-cell therapy where the patient’s own immune cells are collected and modified with viral vectors in a laboratory before being reintroduced to the patient where they can exert their effect against the target cancer cells in vivo…

Cell Culture Medium Development and Analysis for Bioproduction

Over the past two decades, the protein biologics market has exploded and now comprises the fastest growing segment in therapeutics. Chinese hamster cells (CHO) have become the host cell line of choice for the majority of commercial production of protein therapeutics, such as monoclonal antibodies (mAbs), on the market.¹ Other cell lines such as HEK 293 and Vero also play an important role and they are commonly used to produce non-mAb products such globular proteins, viral vector therapeutics, and vaccines…

Cell Therapies Require Well-defined, Optimized Media to Enable Efficient Transition Between Research, Clinical and Manufacturing Scale Culture

The ongoing success of cell therapies in the clinical setting has put an increased focus on optimizing the manufacturing process for these therapies. In particular, cell expansion for clinical use is critical and requires a highly optimized media appropriate for clinical and commercial manufacturing. Increasingly the industry is moving towards using well-defined media without the addition of serum. In addition, clinical and ultimately commercial manufacturing requires a scalable process. This includes a medium that enables large-scale manufacturing systems, such as microcarrier-based suspension culture platforms. To meet these needs, MilliporeSigma has published an article, which describes the use of their Stemline® XF MSC Medium in conjunction with the 3L Mobius® Stirred Tank Bioreactor to increase yield and functionality. The Stemline® XF MSC Medium was designed for efficient expansion in planar and microcarrier-based culture platforms, easing the transition between research, clinical, and manufacturing scale culture…

Deciphering Continued Process Verification and Automating the Process for Straightforward Implementation

Continued process verification (CPV) is used by biopharmaceutical companies to ensure that the manufacturing process for a therapeutic remains in a continual state of control during the lifecycle of the product. This is to make certain that the final drug product’s strength, quality and purity is maintained for the product’s duration on the market. Both the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) have directed manufacturers to develop a CPV program to collect and analyze product and process data to evaluate the state of their manufacturing controls and to identify any product or process problems. It also provides the opportunity to evaluate and implement process improvements…

Enhancement of transient transfection with serum-free and blood-free transferrin

Methods to enhance transient transfection of adherent cells have identified many different ligands that improve plasmid uptake into cells. Of these ligands, serum-derived transferrin is well-known to improve transfection efficiency of adherent cells. However, to date, a serum-free and blood-free transferrin has not been tested for improvements to transfection efficiency. We have thus created a protocol for the improvement of transfection efficiency and viral titer without the need for blood-derived transferrin. Excess transferrin does not negatively impact transfection, and the incorporation of a blood- and serum-free, recombinant human transferrin supports high transfection efficiency and viral titer in a chemically defined, blood-free medium…

Evolution of Viral Vector Analytics for Gene Therapy Manufacturing

With over 350 cell and gene therapies in clinical trials in the US and more than 700 globally, the explosive growth in the field is only expected to increase in the years to come. This rapid progress from clinical development towards product licensure has resulted in a viral vector supply bottleneck. The challenge in viral vector-manufacturing capacity is estimated to be 1–2 orders of magnitude lower than what is needed to support current and future commercial supply requirements. Concomitantly, regulatory scrutiny and product characterization requirements are increasing, as more gene therapy products reach commercialization…

In-Process Analytics are Key to Successful Cell Therapy Process Optimization and Manufacturing

Cellular therapies are an increasingly viable therapeutic option for many indications, particularly for patients who have exhausted traditional treatments. Considerable clinical activity and the approval of several autologous, patient-specific therapies has increased industry demand and has also highlighted bottlenecks in manufacturing workflows. These bottlenecks must be addressed to improve manufacturing productivity, safety, and timely delivery to patients. In an autologous setting, each patient’s health status and demographics translates to a unique cell population, which imparts variability to the subsequent cell therapy manufacturing process. This emphasizes the importance of in-process controls in ensuring a consistent drug product, even in the presence of such variability…

Make Time for Process Development in Cell Therapy Production

Methods for growing cells in a dish have advanced at an extraordinary rate over the last 100 years. This ability to grow cells, as the therapeutic agent, has paved the way for advanced cellular therapies. For these therapies, millions, and sometimes billions of cells need to be produced under highly controlled and consistent manufacturing conditions. Given that cells are complex, require a specific growth environment, and can be directly impacted by cell culture conditions, growing cells to therapeutic levels can be extremely challenging. Many methods for culturing cells at small scale do not translate to large-scale bioproduction. This necessitates process development or the optimization and development of small-scale cell culture methods into processes designed for large-scale bioproduction…

Media panels for AAV manufacturing: Finding the formulation for success

Utilizing media panels to select and optimize HEK293 suspension media for AAV production can accelerate the process and drive more gene therapies through clinical evaluation to commercialization. Media panels comprise a range of nutritionally diverse media formulations for manufacturers to test in their process with their cell line, expediting the selection process and allowing them to progress through media optimization much quicker. Media panels are revolutionizing the way AAV manufacturers establish optimized media for their vector manufacturing processes and present several benefits, which will be discussed below…

Microfluidic Transfection Device is Poised to Remove Current Transfection Limitations for Gene Therapy Applications

The transfer of materials into cells via transfection is a critical operation of manipulation and engineering of cells. DNA is often used as the cargo for transfer; however, other cargos may include RNA, proteins, small molecules, and ribonucleoproteins, such as those used in the engineering of cell therapy products^. Even though cell transfection was initially developed decades ago, challenges remain with the process. Current methods may damage cells, require special reagents, that may be mixing and time dependent, and often inefficient. Performance may suffer since many approaches rely on the passive diffusion of material into cells, usually though temporarily, induced holes in the cell membrane. These challenges, in turn, create burdens in many areas of current biomanufacturing such as manufacturing at scale or in the development of complex, multiple gene-edited cell-based therapeutics…

mRNA Vaccines: Current Trends and Perspectives

The COVID-19 pandemic brought mRNA vaccines to the spotlight with the rapid release of highly efficacious (94-95%) vaccines by Pfizer/BioNTech and Moderna.  Once the sequence of SARS-CoV-2 was published, Moderna had its first candidates available in only 28 days. Full phase 1-3 trials and release of millions of doses were completed in months – much shorter than the years of time typically required for other vaccines. Besides shorted development time and high efficacy (at least for COVID), there are other advantages for the use of mRNA for both prophylactic and therapeutic vaccines. One is the safety profile, which includes that the antigen is typically expressed for only a matter of days and can be modulated by the design of the mRNA.  mRNA vaccines are also much more controllable than attenuated and vector vaccines.  Unlike DNA-based approaches, mRNA vaccines do not require nuclear entry so there is less risk of genomic integration and mutagenesis.  Lastly, mRNA vaccines offer the robust development of both cellular and antibody responses, which can be somewhat shifted between the two by the design of the mRNA, the choice of delivery method, or other approaches…

mRNA Vaccine Production and Facility Design

Over the past year we’ve witnessed an explosion of activity in both vaccine research and manufacturing due to SARS-CoV-2. We’ve also seen an increased appreciation of what had been some rather obscure, though highly significant, technologies. These include vaccine types and sub-types, ranging from exosome encapsulated AAV vectored vaccines in a prime-boost system, to the lipid nanoparticle delivered mRNA of a viral antigen. While traditional platforms remain in use, innovative delivery systems now include entirely synthetic antigenic structures, sub-viral particles, and chimeric bacterial ghosts. Antigen production platforms employ such diverse approaches as animal tissues, cell-free expression, whole plants, cell culture and direct chemical synthesis…

The need of a therapeutic gene editor to develop the next generation of CAR T Therapy

Two chimeric antigen receptor (CAR) T-cell therapies (Kymriah^® and Yescarta^®) are approved by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) to treat B-cell acute lymphoblastic leukemia in children and young adults and diffuse large B cell lymphoma in adults, respectively. Both therapies resulted in high objective response rates (~80% of treated patients) during clinical trials  (Neelapu, 2017) (O’Leary, 2018). Despite these approvals, questions remain over whether cell therapies, such as CAR-T-cells, represent a new era of frontline therapy with potential significance to match that of monoclonal antibodies or whether they will be an effective but niche approach for a narrow range of indications and a relatively small number of patients. This is primarily owing to the challenges encountered when treating patients with solid as opposed to liquid tumors. It is likely that CAR T-cells will need to up their game through genetic engineering if they are to have efficacy in the notoriously immunosuppressive solid tumor microenvironment…

Next Generation Proteomics is the Missing Piece in Completing the Precision Medicine Puzzle

Twenty years ago, the International Human Genome Sequencing Consortium constructed a map of the human genome and with it came the opportunity to look at medicine in new way. The idea of precision medicine, an approach to medicine where an individual’s genes, environment, and lifestyle is considered in disease treatment selection, was enabled for the first time and with it the hope for targeted therapeutics. In spite of this significant advancement, we still primarily employ a one-size-fits-all therapeutic approach to medicine today. Unfortunately this approach does not yield the kind of results that precision medicine aspires to. Why have we not been able to fully employ precision medicine? Many believe that this is due in large part to the inability to harness proteomics as the missing piece of the precision medicine puzzle…

Optimization of conditions to support efficient HEK293 cell growth and adenovirus production

Viral vector-based vaccines have been a research focus for decades leading up to the recent approval of the vesicular stomatitis virus (VSV) vector-based and human adenovirus-based Ebola vaccines. The SARS-CoV-2 pandemic has further highlighted the continued need for vaccines against emerging and re-emerging pathogens. In response, global vaccine manufacturers have developed several types of effective SARS-CoV-2 vaccines, including adenovirus (AdV) vector-based vaccines. However, the production of large-scale, high-titer viral vector vaccines requires optimization of several cell culture parameters…

Publication review: Formulation and production of a blood‐free and chemically defined virus production media for Vero cells

Developing serum-free medium candidates for Vero (African monkey kidney epithelial) cell culture is both a topical and significant breakthrough, as there is an increasing intersection between vaccine development, raw material availability, and regulatory pressures for enhanced quality and safety. Leveraging recombinant proteins in cell culture has garnered increasing interest as a means to enhance the safety and consistency of virus production during manufacturing leading to improved virus-based vaccines and therapeutics…

Recombinant Human Serum Albumin Excipient – Regulatory Filing and Approval for Use in Vaccine Manufacturing

During the 2020 Excipient World Conference & Expo, supported by IPEC-Americas, Kara Quinn, the Associate Director of Engineering for Merck & Co, presented a webinar detailing the first-hand account of the complex process of regulatory agency filing that led to the regulatory approval, by both the FDA and EMA, for the use of a novel excipient, Exbumin™, in the manufacture of their first-in-class Ebola vaccine ERVEBO®…

Selecting the Appropriate Scaling Strategy for Different Cell Therapy Applications

Cell therapies represent a new paradigm in medicine, and the commercialization of several treatments has prompted a need for those developing these therapeutics to look at the scalability of their manufacturing platforms. The decision on how to scale-up and scale-out while choosing the appropriate platform and associated technology relies heavily on the therapeutic being manufactured. Because the cells are the therapeutic product, they need to retain their phenotype and functionality regardless of the manufacturing method. Utilization of primary cells to develop these therapeutics presents unique challenges. These include addressing issues such as donor variability, increased sensitivity to changes in their environment and downstream considerations for formulation and final fill…

Upstream Manufacturing of Gene Therapy Viral Vectors

The majority of gene therapy applications in development utilize viral vectors to carry the therapeutic gene into the target cells. Cells may be genetically modified either in vivo or ex vivo. In ex vivo applications, cells are modified in culture, which also allows for cell expansion and analytical characterization prior to re-infusion of the treated cells. This process has an advantage in that cells can be examined pre-treatment and post-treatment for viability, density, expression level, etc. The ex vivo process can sometimes result in more efficient transduction due to the greater control of the conditions. Alternatively, cellular modification by vectors can also be performed in vivo-directly in the subject. This approach avoids the need for cellular implantation…

Utilizing Optimized Protein Expression Systems to Advance Development of Therapeutics Against Infectious Diseases

The rapid emergence and spread of novel infectious diseases such as COVID-19 caused by the SARS-CoV-2 virus has highlighted the need for innovative protein expression solutions to support rapid and comprehensive virus characterization, serological testing, therapeutic antibody and vaccine development to mitigate these threats to global health. A recent white paper published by Thermo Fisher Scientific entitled Advancing the development of therapeutics against infectious diseases with optimized protein expression systems highlights how their comprehensive portfolio of mammalian and insect transient protein expression systems can accelerate and streamline infectious disease research and meet high-yield bioproduction requirements for the development of diagnostics and therapeutics that can mitigate virus spread…