Don’t miss our top articles of 2019! I have compiled a list of our most popular 25 articles for 2019 in alphabetical order.

Assay Automation – A guide to understanding the key drivers and considerations

Assay automation can improve a lab’s efficiency, throughput, data quality and permit scientists more time for non-routine tasks. The level of automation for an assay workflow can be flexible with some companies choosing to automate the entire process from sample handling to analysis. In other cases, only certain steps of the workflow may be selected for automation. Often people have the perception that complex assays benefit the most from automation, however there are good reasons to automate basic, routine assay formats as well. In this article, we will look at the benefits of automating assay workflows and also the key considerations important to deciding whether automation is right for your process…

Biologic Products DNA to IND Timeline in 9 Months – Yes it can be done!

The ability to rapidly develop biologic products from conception to human clinical trials is an increasingly important aspect of controlling drug development costs and in expediting the drug’s pathway to one day provide critical treatments. Hence, there has been an increased focus on ways to shorten the timeline through product development. These expedited timelines are especially important when dealing with infectious disease control…

Chemically modified amino acids enable the evolution of fed-batch processes from multiple feeds to single feed strategies

Cell culture optimization is critical to good biomanufacturing. In particular fed-batch cultures require optimization of feed strategies to ensure that cultures are not becoming nutrient depleted, to extend the growth phase and increase product concentration. There are several growth factors, amino acids and supplements that are used in cell culture feeds. While amino acids are one of the most popular choices, they can also be challenging because they are difficult to dissolve at high concentrations. This low solubility particularly in high concentrations necessitates that these amino acids be solubilized in separate feeds at basic pH. This can cause pH spikes, precipitation and early death when added to the bioreactor. One of the most challenging amino acids has been L-Tyrosine…

Clarification and Sterile Filtration of High Cell Density Cultures in One Step

Lab-scale cell removal for discovery and process development applications has traditionally been done by centrifuging the broth and then filtering the sample through a sterile filter unit. This has worked well, but cell densities have increased dramatically to boost productivity and this has created challenges in harvest. High cell density cultures require more time for centrifugation and typically multiple filters need to be used during sterile filtration due to clogging. The inefficiency in clarification and sterile filtration caused by high cell density cultures results in significantly longer filtration times, frequently over 60 minutes…

End-to-End Integrated Manufacturing Solutions Enable Gene Therapy Commercialization

The past decade has seen a surge in the development of gene therapies. Currently there are over 1,000 cell and gene therapy clinical trials underway worldwide.¹ While the majority of current trials are still in Phase I or Phase II, there are more than 90 trials in Phase III. In fact, at last year’s BIO International Convention, Scott Gottlieb, Commissioner of the US Food and Drug Administration (FDA) stated that he expects the FDA to approve 40 gene therapies by 2022…

New Flexible Bispecific Antibody Format Demonstrates Improved Therapeutic Properties and Manufacturability

This year’s Next Generation Protein Therapeutics & Bioconjugates™ Summit held in San Francisco provided an in-depth look at the latest developments in next-generation protein therapeutic discovery and manufacturing. One talk that I found of particular interest was, “WuXiBody,™ an Innovative and Versatile Bispecific Antibody Format, Opens a New Era For Therapeutic Antibody Development,” presented by George Wang, Ph.D., MBA, Vice President of Biologics Discovery, WuXi Biologics. The talk provided an excellent overview of the promise of bispecific antibodies as well as the associated challenges. Dr. Wang also shared an innovative bispecific antibody platform designed to address many current platform shortcomings…

How to get the most out of automating your ELISA-like assays, dos and don’ts

Bringing automation on any scale into your lab can be an intimidating prospect, especially for new users. There can be significant benefits to automating a method or process, but there are also many factors to consider and even more potential pitfalls. It is a common misconception that automation or integrated platforms have to be big, expensive and complex. There is also a common misconception that less complex assay formats don’t benefit from automation. Implementing automation can be as complex as large, walk-away high-throughput screening platforms or as simple as a plate reader and stacker, or any combination in between. Thoughtful planning and careful implementation are the two factors that determine the overall success of the process. Significant care should be taken in properly defining the scope and scale of any automation project. What do you hope to accomplish? Is it increased throughput? More consistency across assay runs? Allow scientists to focus on non-routine tasks? Automation can help provide all of these. Automating relatively simple assays that need to be done on a regular basis is a great way to free up time for scientists to focus on non-routine work while maintaining consistency across assays…

Implementing Digital Biomanufacturing in Process Development

At this year’s Bioprocess International Conference in Boston, Dr. Thomas Seewoester, Amgen gave a very insightful and inspiring keynote presentation titled, “Bioprocessing 4.0 – Digital technologies are transforming biologics manufacturing.” In the talk, he discussed the evolution of biomanufacturing and how digital technologies were a large part of the next step forward. He reminded us that each company is moving at its own pace and that the level of digital implementation is varied across the industry. He encouraged us as an industry to think outside the box and to look at how other industries are incorporating digital technologies to see what we can apply to biomanufacturing…

Increased CHO Transfection Efficiency and Production in a Straight Forward System

Transient protein production enables quick and efficient production of milligram to gram quantities of recombinant protein, which saves time and cost compared to developing a genetically stable cell line for use in bioproduction. This is particularly important in drug development where preclinical material is needed quickly in order to make informed go/no go decisions. Having access to preclinical material faster can greatly impact the overall drug development timeline. The challenge has been low transfection efficiency and productivity, particularly in CHO cells. CHO cells are typically more difficult to transfect, but are the preferred vehicle of antibody production due to their use in larger scale production of clinical material…

Innovative Chemicals for Process Intensification in Cell Culture Media

L-Tyrosine is a key amino acid for both cellular metabolism and protein synthesis and its depletion in fed-batch processes has been correlated with a drop in specific productivity¹ and with protein sequence variants.² This critical amino acid presents an extremely low solubility, especially at neutral pH.^3,4 The use of tyrosine di-sodium salt concentrations above 1 g/L in feeds induces precipitation and increases the risk of media instability, mainly through co-precipitation of other amino acids. This may lead to sub-optimal performance due to insufficient supply of nutrients and finally to low performing processes…

In-Process Analytical Methods are Critical in Gene Therapy Development

In gene therapy manufacturing, evaluating viral vector stability and integrity is critical in process development prior to production scale up. Process changes can affect critical quality attributes and as a result any process changes need to be carefully evaluated to determine the impact on viral particle size and intactness to ensure final product quality…

Intelligent Lentivirus Analysis – Coupling an Automated Imaging System with Artificial Intelligence

The number of gene and cell therapies in the clinic is increasing rapidly. With the increase in gene therapy manufacturing, comes the need for improved analytics during the production process. Analytics for Lentivirus production is particularly important as it is currently the second most common vector used in cell and gene therapy manufacturing…

Keys to successful single-use bioreactor scale up – from small scale to pilot scale

Successful monoclonal antibody (mAb) manufacturing relies on good process development and reliable bioreactor scale up. Initial process development work is typically conducted at small scale, then moved to bench scale and lastly to pilot scale before moving into manufacturing. One of the biggest challenges is scaling up an optimized process from process development to manufacturing scale. Just a small change in the size of the bioreactor can have a cascade effect on many other culture conditions…

Minimize Cell Culture Contamination and Batch Failure by Creating a Closed System for Cell Culture Expansion

Batch failure due to cell culture contamination is a serious consideration for biomanufacturing. A recent BioPlan Associates Survey examined the frequency, cost and leading causes of batch failure. They found that on average a failure occurs every 9.4 months and costs companies $1 to $2 million. The leading causes of batch failure were contamination and operator error. It is logical that anytime the cell culture system is open, for instance when opening flasks during cell expansion, there is an opportunity for contamination…

A New Dimension of Cell Culture: The Rise of Spheroid Culture Systems

In this article, we discuss the evolution of spheroid culture systems, methodologies of spheroid formation, spheroid culture challenges and relevant solutions. Historically, much of the knowledge we have on cellular processes is from experiments where cells cultured on flat, two-dimensional (2D) surfaces. 2D cell culture systems span many applications from studying basic cellular processes, stem cell and cancer research to regenerative medicine. While these systems are attractive because they are simple, cheap, and work for a variety of cell types, there are some limitations. Cells cultured on planar surfaces do not reflect the in vivo physiology such as tissue-specific architecture, cell-to-cell and cell-to-matrix connections, which can affect their proliferation, function and response to external stimuli¹. This is evidenced by the fact that 2D model systems are not always predictive, resulting in disappointingly high failure rates when used in new drug discovery and clinical trials¹. Despite these disadvantages, 2D cell cultures are still routinely used because they are well-established and historical data based on 2D cultures allows researchers to execute comparative studies. Additionally, most of the equipment used for high throughput screening (HTS) is compatible with and optimized for 2D culture systems making them amenable to large scale screening protocols…

Optimising outgrowth of clones following single cell cloning ensures more predictable and effective cell line development

Successful cell line development is critical to effective CHO cell based biomanufacturing. To accomplish this, companies must identify cell lines that have good manufacturing potential via clone selection. Clones that warrant further investigation are those that are stable, good producers, demonstrating specific attributes based on the product specifications. In order to select the best cell lines, companies must generate a good number of clones for screening…

Qualifying a rapid qPCR mycoplasma test approved for CAR-T clinical studies

Like other biologics, most cell therapy products require microbial testing including, mycoplasma, sterility, and endotoxin analysis as part of the final product release criteria. However, cell therapy products have unique QC product release requirements that necessitate new microbial testing technologies to support these unique needs. One of these is that since the cells are the final product so they cannot be put through a final sterile filtration step or undergo a harsh sterilization step and have a limited shelf life. As a result, it is a critical to implement rapid microbial testing for in-process and final product release that are compliant with the stringent regulatory requirements and also meets the unique QC needs for cell therapy products. Traditional testing methods for mycoplasma, sterility and endotoxin can take several weeks to over a month and require a large amount of sample volume which doesn’t fit the needs of limited sample availability and quick turnaround time for cell therapies. This is why rapid microbial methods, including rapid PCR mycoplasma methods, provide an important solution to a significant challenge in cell therapy quality management…

Rapid Mycoplasma Testing for Biomanufacturing Applications

Increasingly the quality of raw materials used in cell culture media and supplements is being closely examined for any potential negative impact on cell culture or the final product quality. One area of concern is biomanufacturing contamination. While there are certainly many possible entry points for cell culture contamination, contaminated raw materials is high on the list. Of the possible contaminants, mycoplasma is often a big concern because it is fairly resilient in laboratory environments, resistant to most, if not all antibiotics, and also contamination in culture is not always obvious. Yet, mycoplasma contamination can have a devastating impact on biopharmaceutical development and production. As a result, having adequate mycoplasma monitoring in place is critical for minimizing the risk and ultimately impact of contamination…

Scaling Strategy for Moving mAb Process from Microscale to Production Scale Bioreactors

The most efficient process optimization requires successful scale-up and scale-down of manufacturing processes. Scaling strategies are key to ensuring that the scaled-down process will scale-up as expected and that the scaled-down process is a true representation of larger scale manufacturing. A strategic scaling approach is important to determine the bioreactor operating parameters to be kept constant during scaling. While critically important, a good scaling strategy is not a simple task and I am pleased to share the following guest blog that walks through creating a scaling strategy for moving from microscale bioreactors to production bioreactors. I was fortunate to be able to interview the author about his article and have provided the transcript of our conversation following the guest blog…

Spheroid Cell Culture – Practical solutions for frequently asked questions

The past decade has seen an increase in publications recognizing the value of culturing cells in three dimensions (3D) and the use of these 3D cell culture models has shown utility in many areas of research from cancer biology to regenerative medicine. One of the most common 3D cell culture types is spheroids. Spheroids more accurately recapitulate the native in vivo microenvironment with respect to cellular function and response to drugs than more traditional two-dimensional (2D) models. That said, working with spheroids is not without its challenges. Many labs working with spheroids have run into technical hurdles incorporating them into existing cell culture workflows. Whether it is issues with handling or optimizing protocols to assaying cells in 3D, these obstacles can be overcome…

Spheroid Culture Advancements – Microplates

Spheroid cultures are on the rise as the success of 3D spheroids has been demonstrated in many areas of research including anti-cancer, drug screening, and in vitro tumor studies. With increase in use, has come the need for better tools to enable the culture of these spheroids with uniform size and in large quantities. These methods must also be repeatable and efficient…

There’s a New CAR in Town: CAR-Expressing Natural Killer Cells

The ability of the body’s immune cells to target and eliminate infectious organisms and foreign invaders has been studied for decades. It is no surprise that scientists want to harness the power of these cells in immunotherapy to specifically target and eliminate unwanted cells in the body. CAR (chimeric antigen receptor)-expressing T cell adoptive cell therapies have generated a lot of excitement and investment in recent years due to the unprecedented clinical results, which is driving research forward at a breakneck pace. The general steps for CAR-T are as follows: T cells are extracted from a patient’s blood, genetically modified with a chimeric antigen receptor specific to a surface antigen on target cells, culture-expanded and then reinfused back into the patient. To date, the U.S. Food and Drug Administration (FDA) has approved two CAR-T therapies (Novartis’ Kymriah® and Kite/Gilead’s Yescarta®) targeted against hematologic malignancies, and currently hundreds more clinical trials are underway for other malignancies, including solid tumors. However, the autologous (patient-specific) nature of this cell therapy, complex manufacturing workflows and the risk of graft versus host disease (GVHD) has raised concerns over its cost and safety. Similarly, obtaining enough lymphocytes from an ill, often lympho-depleted patient can pose a barrier to generating clinically relevant doses of CAR-T cells…

Transfection Reagent Portfolio Enables Seamless Scalability from R&D to Commercial Therapeutic Viral Vector Manufacturing

Cell and gene therapies are achieving greater success in the clinic and as a result there is an increasing need to transition manufacturing from research scale to clinical and ultimately to commercialization. Critical to the manufacture of many gene-based therapies is the production of large quantities of recombinant viral vectors. Two commonly used recombinant viral vectors, Adeno-Associated Virus (AAV) and lentivirus, are mainly produced using PEI-mediated transient transfection in adherent and suspension HEK-293 or HEK-293T cells…

Using Organoids for Disease Modeling

The culture of organoids has permitted researchers access to a highly physiologically relevant system for studying human disease. Human organoids reflect key structural and functional properties of organs such as kidney, lung, gut, brain and retina making them incredibly valuable, particularly in situations where there is an unmet need for an in vitro human model. While their value is undeniable, culturing organoids can be quite challenging. Many tools and techniques have been developed for organoid culture, however depending on the cell type, research area and experimental goals, it can be difficult to identify the right method…

Using transposases to generate clones with high, predictable expression and stability

Standard cell line development workflows include transfection followed by the screening of hundreds to thousands of clones, in an effort to find high producing clonal colonies that are stable and demonstrate specific attributes based on the product specifications. Some approaches at the clone isolation stage utilize in-situ fluorescence detection to try and pull out up front the best protein secretors…