Rank #1: Bi-Specific Antibodies – The development, manufacture and promise of these cutting-edge therapeutics
In this podcast, we conducted a panel discussion with experts from Selexis and KBI Biopharma on bi-specific antibodies. We examined bi-specific antibody development and manufacturing, including current challenges and key solutions. We also discussed the promise of these cutting-edge therapeutics and their future in medicine.
May 26 2020
Rank #2: New Advancements in Cell and Gene Therapy Implementing Precision Medicine
In this podcast, we talked with Dr. Paul Wotton, Chief Executive Officer, Obsidian Therapeutics about the evolution of cancer therapy and new advancements in the space including precision medicine. Our discussion included Obsidian’s cytoDRiVE platform that provides a technology, in which the level and timing of protein activity are fully controlled in a dose-dependent manner by an FDA-approved small molecule.
Dec 05 2019
Rank #3: Viral Vector Manufacturing for Gene Therapy
In this podcast, we interviewed Dr. Mats Lundgren, Ph.D. Customer Applications Director, Life Sciences, GE Healthcare, Sweden about developing platform processes for viral vector manufacturing and Mats describes a recently developed platform process for adenovirus production.
Jan 16 2019
Rank #4: Expediting cell and gene therapy workflows
In this podcast, we interviewed Mr. Dave Eansor President of the Protein Sciences Segment at Bio-Techne and Dr. Sean Kevlahan, Senior Director of Cell and Gene Therapy at Bio-Techne about new technologies that can be implemented to expedite cell and gene therapy workflows and facilitate the path from bench to clinic.
View show notes at cellculturedish.com/expediting-cell-and-gene-therapy-workflows/
Feb 06 2019
Rank #5: Improved gene therapy analytics permit monitoring of critical quality attributes and increased manufacturing efficiency
Jonathan Royce, Director, Instruments Business Unit, Vironova talks about analytics in gene therapy manufacturing. Specifically, Jonathan details new technologies to monitor critical quality attributes, reduce development time and increase the number of projects that can be run.
Dec 10 2019
Rank #6: FBS – It’s Not All Created Equal – What Consumers Need to Know
In this podcast, we interviewed Alyssa Master, Ph.D., Senior Manager of Science and Applications, Nucleus Biologics about some of the challenges of working with FBS, why not all FBS is created equal and suggestions for FBS customers who are trying to navigate these issues.
We began the interview by discussing what FBS is and how it differs from other types of serum. Alyssa explained that FBS is a bi-product of the meat and dairy industry and it is great for in vitro cell growth because it contains a cocktail of growth factors. FBS is biochemically different from other sera like adult bovine serum or even newborn calf serum. Due to these biochemical differences, FBS has become the gold standard and as a result it is more expensive.
Alyssa continued that because of its ability to support cells in vitro it is widely used throughout academia, biotech, large pharma and bioproduction, particularly in cell therapy and vaccine biomanufacturing. I asked why so many use FBS and Alyssa said that it works well for most lines and is readily available. Because it is animal derived, it already contains cytokines and growth factors in the right combination to support cell growth.
View all show notes at cellculturedish.com/fbs-not-created-equal-consumers-need-know/
Nov 08 2017
Rank #7: Key Considerations in Gene Therapy Manufacturing for Commercialization – A panel discussion
This podcast is a recording from the panel discussion that I moderated at this year’s Cell and Gene Therapy Bioprocessing and Commercialization Conference. The discussion covers the latest in gene therapy manufacturing strategies, analytical analysis, cost of goods and key regulatory considerations.
Dec 05 2018
Rank #8: Innovative vaccine manufacturing enables the delivery of vaccines to the developing world
In this podcast, we interviewed Dr. Alex Chatel, Product Manager, Viral Applications, Univercells about the biggest challenges facing vaccine manufacturing today, why it is so difficult to manufacture and deliver vaccines to the developing world and how a novel technology with the support of the Gates Foundation is poised to address these challenges.
I began the interview by asking Dr. Chatel what he sees as the biggest challenges facing vaccine manufacturing today. He stated that the main challenge is manufacturing capacity and as a result, vaccine supply is too low compared to what is needed. Another big problem is that vaccines are being manufactured today using technologies that are outdated. These inefficient vaccine manufacturing methods cause both captial expenditure and operating expenses to be higher than necessary. Unfortunately the cost of production of vaccines will remain too high unless a change in manufacturing is implemented.
Next, I asked him why it is difficult to manufacture and distribute vaccines to the developing world? He explained that the capacity and outdated technology problems are issues in both the developed and developing world. However, the developing world has the additional challenge of distribution, specifically cold chain. Maintaining product stability is obviously important to maintaining efficacy, but cold chain distribution is often hard to maintain from manufacture to delivery in some of these areas. Some companies are working on developing more stable formulations for travel and other solutions. Administration of the vaccine and managing the proper handling and disposal of syringes is also difficult and presents the risk of spreading infectious disease.
We then discussed how Univercells’ Scale-X technology helps enable vaccines delivery worldwide? He explained that most viral vaccines are manufactured using adherent cells and thus they need a point of anchorage for production. Currently support matrices used in vaccine manufacturing include microcarriers in bioreactors or static systems such as roller bottles or stacked plate equipment. Scale-X couples the advantage of having the environmental culture control found in bioreactors with the gentle growth environment of static systems. In this system, the cells adhere to the support matrix, while nutrients in the culture media are circulated around the cells gently. On of the key advantages of this approach is that a high level of production can be achieved in a small footprint of equipment. This enables better facility design and utilization, plus capital expenditures are much less than traditional manufacturing technologies where more facility space and environmental controls are necessary.
Another advantage that Alex shared is that the equipment is small and fully automated with only a small number of operations required to run the system. This provides a much less risky and less costly alternative to traditional manufacturing where more manual operations increase risk of error and require more labor to operate. In addition, the Scale-X technology is highly scalable and permits easy and quick scale up to larger volumes. Cells receive a consistent experience at both small and large scales, so scale up is quite simple.
Alex then explained how Univercells has demonstrate success of the platform through hundreds of experiments using vero cells to produce polio vaccine. Through a grant given by the Bill and Melinda Gates foundation, they have looked at improving the manufacturing of the polio vaccine from R&D to pilot scale and are now ready to scale up to large scale production. They plan to manufacture their first lot of clinical material and have also been gathering experience with other cell types and viruses for vaccine manufacturing and the manufacturing of viral vectors for cell and gene therapy applications.
I then asked if Alex could tell me a little more about their work with the Gates Foundation. He said that one of the goals of the Gates Foundation is a focus on eradication of polio from the planet and they fund programs to achieve this goal. One area of their funding is aimed at disruptive manufacturing technologies, which Scale-X represents. The goal here being to bring new technologies to vaccine manufacturing that will drive down the cost of vaccine manufacturing and furthermore make it more commercially interesting for companies to develop and produce new vaccines. Also, there is a need to fill the current supply gap for existing vaccines of interest in order to reach worldwide vaccination targets. Univercells received their grant in 2016 and will have the first lot of clinical material with the ultimate goal of manufacturing polio vaccine at large scale.
I went on to ask Alex how he envisions Scale-X being employed in the developing world. He said one of the key features of the technology is its ability to integrate with another product to create a micro facility. In this micro facility both cell culture production of viral material and also purification could occur within a small environment. This would cause a drastic reduction in required manufacturing footprint and the entire system could be quickly installed in countries where there might not be the existing manufacturing structure that exists in the developed world. The main barrier to entry for developing world that wants to manufacture its own vaccines is the cost of building a manufacturing facility. With the micro facility, these countries could consider manufacturing their own vaccines rather than purchasing from pharmaceutical companies.
Last, I asked Alex if he had anything to add for listeners. He said that one of the cool things about this technology is that it represents a step change in how vaccines are manufactured now and in the future. Once implemented it would be completely different to what we are used to and presents the opportunity to bring more modern technologies to vaccine manufacturing -an industrial revolution of vaccine manufacturing. Ultimately though the main benefit is that disease eradication targets through vaccination can be more easily met.
Jun 11 2019
Rank #9: Antibody Drug Conjugate (ADC) Development and Manufacturing Challenges and Solutions
In this podcast, we interviewed Dr. J.J. Luo, Executive Director and Dr. Lily Yin, Head of Biologics Conjugation Development, at WuXi Biologics about the state of the ADC marketplace and the challenges in developing antibody drug conjugates. We then dove into reasons behind why they built a dedicated manufacturing facility for ADCs and other bioconjugates and how a single-source development platform can benefit bioconjugate drug development efforts.
I began the podcast by asking Dr. Luo about WuXi Biologics’ investment in and construction of multiple clinical and commercial scale facilities around the world. I pointed out that nearly all of these facilities are dedicated to the GMP production of Drug Substance and Drug Product of antibody or recombinant protein therapeutics, but their new facility appears to be dedicated for a very specific niche within the biologics industry. JJ explained that the new facility was purpose-built for the manufacture of a wide range of bioconjugates, in which the largest class of these products would be Antibody Drug Conjugates. He went on to say that this new facility is part of WuXi Biologics’ new state-of-the-art integrated Biologics Conjugation Solution Center in Wuxi City, China. The center will consolidate all bioconjugate drug development and commercial manufacturing of bioconjugates in the future.
I followed up by asking JJ about the drivers behind building a facility dedicated solely to ADCs and other bioconjugates. He stated that although bioconjugates are a subset of the greater biological therapeutic marketplace, various market research reports have shown a year on year increase in the number of ADCs and other bioconjugates entering clinical trials over the past 5 years by roughly 12% per year worldwide. Some estimate that commercial sales of ADCs will grow 22% annually for the next 5 to 10 years. In addition, based on drug company pipeline data, it appears that this trend will continue into the foreseeable future. JJ made it clear that WuXi Biologics’ mission is to provide open access drug development platforms to ensure that every drug can be made and every disease can be treated. Given WuXi Biologics’ expertise, vast capabilities, and resources they believe that they can provide a single-source product development and supply chain for companies developing these novel therapeutics.
Dr. Yin added that WuXi Biologics has a strong history of providing extensive product development and manufacturing services to clients. They have developed a wide variety of bioconjugates from idea to GMP manufacturing for clinical study. She goes on to share that they have provided full CMC services for 11 IND filings for bioconjugate products in just the last few years and 25% of the total IND filings for ADCs in 2018. With the new bioconjugation facility, WuXi Biologics’ has increased GMP manufacturing capacity for both drug substance and drug product and can meet their client’s needs for clinical supply while still offering additional capacity for new clients.
Next, I asked Dr. Yin about their ability to provide a single-source for all bioconjugate needs. I clarified that ADCs and other bioconjugates are complex molecules with both chemical and biological components. I wanted to know how they would be able to provide all the services required to get from early discovery to clinical supply and if there were any gaps. Lily said that WuXi Biologics is known for their ability to take clients from early discovery and lead generation of antibody therapeutics into development and the clinic. Through WuXi Biologics’ expertise and single-source services they are able to provide this without having to use another service vendor. She went on to say WuXi Biologics is now a separate company, but it is still a member of the WuXi AppTec Group of companies. They still work very closely with the Group and utilize its vast resources and capabilities in discovery chemistry, development and GMP manufacture of small molecule payloads and linkers. In addition, WuXi Biologics, along with WuXi AppTec, can provide the essential preclinical services such as DMPK, oncology research, early stage IND-enabling toxicology amongst other critical services. Couple these single-source services with the fact all of these services and the entire supply chain are within minutes to a couple hours from each other and WuXi Biologics can provide a significant advantage in time and also risk reduction for any company wanting to develop these type of drugs. Lily encouraged listeners to go to their website to learn more about their comprehensive capabilities. https://www.wuxibiologics.com/services-solutions/antibody-drug-conjugates/
Then we discussed how some of the new bioconjugate modalities often contain highly potent or highly toxic payloads thus requiring development and manufacture of these intermediates and the subsequent conjugation to be done in special facilities equipped to handle these compounds. They often also require additional oversight from regulatory authorities. I asked JJ how WuXi Biologics is addressing these types of intermediates and compounds. JJ described how their new facility has been purpose-built to handle highly potent and toxic materials. The design and construction were set to meet U.S. FDA, EMA and China National Medical Products Administration (NMPA) standards for those compounds.
We then focused on how ADCs have historically had many challenges in development and also challenges in demonstrating efficacy in humans. I asked Lily what is the industry doing to help mitigate these development challenges, reduce risks, and help ensure success to move more of these treatments into the clinic and hopefully to commercialization. Lily acknowledged that there have been set-backs and challenges in the field, but stated that bioconjugates are still a relatively young field with a lot left to learn on how we can more effectively develop successful drugs. She went on to say that the good news is that there are 5 ADCs on the market with approximately another 10 in late stage clinical trials with over half of those having obtained “Fast-track” status. WuXi Biologics believes that this demonstrates that the industry is beginning to turn the corner with better understanding of how to develop these molecules for clinical success. She shared that there is a better understanding of how to engineer antibodies to find suitable targets, how to select an active toxin, and how to use better linker chemistry to create better targeted binding characteristics, improved in vivo half-life, more consistently controlled drug-to-antibody ratio, residual free drug levels and better payload release to control drug levels in vivo.
I then asked Lily what specifically WuXi Biologics is doing to help further the ADC field. She said that they have now worked with so many different antibody or other biological molecules, linker and payload chemistries and combinations thereof, that they are uniquely qualified to advise clients on development strategies that will best suit their needs and ensure success. They have strong manufacturability and developability strategies when evaluating the various technologies and bioconjugation options to provide sound data driven decisions for lead selection and throughout development in order to generate a robust GMP process. Looking at many of the challenges and pitfalls of certain linker and bioconjugation schemes WuXi Biologics has continually developed their own new technology platforms for linkers and in some cases proprietary modifications to enhance certain payloads to increase their effectiveness for first in human trials.
I followed up by asking Lily to elaborate on the proprietary technologies she mentioned and also to discuss the benefits of WuXi Biologics’ IP. Lily said that they have developed a novel linker for Lysine-based conjugation that demonstrates higher reactivity, better solubility and a more flexible range of conjugation temperatures and the ability to conjugate this linker to other molecules beyond IgGs. Their unique payload chemistry involves more homogenous drug loading for cysteine conjugation. She went on to say that they would be happy to discuss more under a CDA with any company who is interested.
Next I asked JJ if these technologies were scale-able. He clarified that WuXi Biologics designed all of these technologies with an eye for ease of formulation and to be able to scale up for clinical trial scale and beyond.
I followed up with JJ and asked what manufacturing scales could be achieved at the new facility. He said that the new facility could produce up to 50 batches per year with up to 2 kilograms per batch for drug substance. He said that they can perform both liquid and lyophilization for Drug Product fills with a maximum of 100,000 2R vials per batch for liquid fills and 25,000 2R vials per batch for lyophilized fills. In addition, they are able to provide GMP lyophilization of ADCs in various vial sizes including 2R, 6R, 10R, 20R and 50R. He also mentioned that the facility could perform filling of other non-bioconjugate products such as small molecule or peptide therapeutics as long as they require liquid or lyo type fills. He added that since this is a multi-product and multi-use facility, they are utilizing single-use systems including single-use compounding vessels, tubing, bags, and filling needles for the entire bioconjugation and drug product compounding, filling and finishing process. They do this to greatly reduce risk during the GMP manufacturing runs and provide the highest level of quality assurance for their clients.
I asked that since they are working with highly potent and toxic compounds in this facility were there any special equipment requirements. JJ shared that isolator-based systems are widely utilized in the industry for working with highly potent or highly toxic materials, so they utilize isolator-based systems for payload and linker dispensing and dissolution, filling and lyophilization. In addition, fully automated equipment is used for vial washing and depyrogenation, filling, and for automated loading and unloading the drug product containers into and out of the lyophilizer. Filling is operated by an automated peristaltic pump dosing system and weight check is performed using on-line weighing system for automatic fill weight adjustment. If required, the filling line is also equipped with Nitrogen blanketing capabilities and light protected material handling is also available from start of fill processing to final package steps.
Lastly, I asked when they will start GMP ADC manufacturing activities. JJ said that they have just started operating the facility and their first GMP production will be in the third quarter of this year. There are nearly 20 lots that are scheduled including 10 GMP lots by the end of this year. So, although he said that they will be busy, there is still capacity for this year and next year as well.
I closed by thanking JJ and Lily for their time. The interview was very informative and provided good insight into the current state of the ADC/bioconjugate marketplace and how the industry and WuXi Biologics is working to overcome the historical challenges. It looks like a bright future for bioconjugated therapeutics is ahead of us.
Sep 05 2019
Rank #10: Mesenchymal Stem Cell Culture – Challenges and solutions for isolation, expansion and maintenance
In this podcast, we talked with Dr. Jennifer Chain, Scientific Director of Research and Development, Oklahoma Blood Institute about the isolation of mesenchymal stem cells form cadaveric bone marrow and the differences between live and cadaveric donors. We also discussed the expansion and maintenance of mesenchymal stem cells in culture including media design and selection.
Mar 12 2020
Rank #11: Closed System Cell Therapy Manufacturing – Moving from the lab to clinical and commercial production
In this podcast, we conducted a panel discussion on key considerations for moving cell therapies from lab to commercial manufacturing, including the benefits of manufacturing in a closed system. We also discussed what were some of the key hurdles and possible solutions associated with transitioning to clinical and commercial cell therapy manufacturing.
Jul 26 2018
Rank #12: Using 3D cell-based human liver microtissue models in predicting adverse effects caused by chronic exposure to engineered nanomaterial
In this podcast, we interviewed Shareen Doak is Professor of Genotoxicology and Cancer in Swansea University Medical School and Dr. Wolfgang Moritz, Co-founder and Head of External Collaborations and IP, Insphero about the use of 3D cell-based human liver microtissue models to predict the adverse effects of chronic exposure to nanomaterial . We also discussed the use of liver microtissue models for drug discovery and development.
Oct 30 2019
Rank #13: Successful Mesenchymal Stem Cell Manufacturing for Cell Therapy Applications
In this podcast, we interviewed Dr. Ross Macdonald about the use of mesenchymal stem cells for cell therapy applications and key criteria for successful manufacturing.
May 30 2018
Rank #14: How Live Cell Analysis Technology is Meeting the Needs of Ever Evolving Advanced Cell Models
In this podcast and accompanying article, we interviewed Dr. Kimberly Wicklund, Head of Product Management for the IncuCyte at Sartorius about how live cell analysis is meeting the needs of advanced cell models and how the launch of the new IncuCyte SX1 is providing scientists more options when it comes to using live cell analysis in their workflows.
We began the podcast by discussing how cell modeling and cell systems have evolved significantly. I asked Dr. Wicklund if she could explain for listeners how cell monitoring has advanced to meet these needs. She described how researchers are looking for physiological relevance in their cell models and cell systems and are making a shift from simple, recombinant cell systems, and moving towards primary cells or stem-cell derived cells that are often human and patient-specific. Co-cultures, multi-cultures and tissue organoid models offer significant promise, but also present challenges to the typical cell workflows, which puts huge pressure on instrumentation to keep up. She went on to stress the importance of taking cell health into account first and foremost in instrumentation, but also to consider the end users as well to ensure that their work can be done efficiently and that their cells are being used efficiently as well.
Next I asked Kim if she could explain live cell analysis and how is it different from endpoint workflows like flow cytometry. She explained that live cell analysis centers on the dimension of time and it centers on repeated measurements of the same populations of cells. She then provided a great analogy, she said “If you think about your favorite sport, do you want to see the score half-way through and nothing else, or do you want to watch how it evolved? If you only see the score at a point in time, how do you know who won, and how do you feel about the next game? You would have no idea what’s to come.”
We then discussed why live cell analysis a good fit for advanced cell systems like primary cells and stem cell derived cells. Kim continued with her sports analogy by explaining, “if you’re watching a game, you want to know what team started strong and then lost their edge, what players really shined, that can change in an instant. So think of your cell models as your favorite team, full of different personalities that can be considered unpredictable until you watch them long enough and really dig in and put them in an environment that doesn’t change and then and only can you begin to predict.”
Next I asked Kim if she could get a bit more specific and describe the IncuCyte live-cell analysis technology and why is it a good fit for these more advanced cell models. She explained that it starts with being an advocate for the cells and the scientists. She went on to say that with the IncuCyte, cells are put in a precise and robust environment, a standard tissue culture incubator. Cells are then left in one spot and the IncuCyte’s mobile optical system travels to the cells and captures images repeatedly, over time. For the scientists, the IncuCyte is as simple and as automated as possible from placing the cells into the IncuCyte until the time when you are getting results. She said that during product development, we have our team of biologists that are sitting right alongside the engineers saying, “I need this to be fewer clicks” or “I need this to suit my workflow” or “I don’t want to figure that out, that’s more technology than biology.” And that’s the rule, if it gets too complicated, then we’re wasting cells and we’re wasting scientists’ time. And we don’t like to do that!
We then talked about an exciting announcement that Sartorius is making at ISSCR this week. Kim explained that they are expanding the IncuCyte product portfolio to include another model, the IncuCyte SX1. Kim explained that the SX1 offers the same information-rich analysis and streamlined user experience as the flagship model, the IncuCyte S3. However, she described that the S3 is a workhorse that can accommodate multiple experiments in parallel, up to six microplate experiments at a time, which is ideal for a lab that has a substantial workload with many users performing live-cell analysis on a daily or weekly basis. But not every lab has that demanding of a workload. A lab might be new to live cell, or there may be fewer users, and that is the lab that might prefer an SX1.
I told Kim that scientists will want to know how it will make their research better, more efficient and more productive. She then described some of the key workflow advantages of the IncuCyte including the ability to observe and measure cell models during the entire cell biology workflow, from culture to manipulation to assay and imaging and analyzing cell culture flasks around the clock. She listed several key workflow steps that could be accomplished using the IncuCyte including ensuring cell seeding densities are consistent, validating successful transfection, performing an assay at microplate scale to see what happens in response to a treatment or a knock-down, just to share a few examples. Kim finished by saying that the IncuCyte accommodates a range of applications for analysis of cell health, movement, morphology or function that can easily be adapted to advanced cell models.
I stated that one of the most impressive aspects of the IncuCyte technology to me is that there are over 2,500 peer-reviewed articles discussing applications of the IncuCyte and new uses of are being developed all the time. I asked her how they handle questions from users during implementation of some of these new applications and what kind of support is offered to end users. Kim shared that she thinks one of the greatest strengths of IncuCyte support is the fact that applications are developed from start to finish in their own labs. They take questions from users, sit with biologists, and then provide all the tools that they need from start to finish to answer that question. That means that they develop appropriate reagents, vesselware, and software to get a scientist from point A to point B with as little trial and error as possible, plus they look at multiple and relevant cell types and cell models to evolve applications.
I followed up by asking if they utilized the information from users about new applications in developing the new IncuCyte model. She said yes, what they saw was that every cell biologist had a need to observe their cells across the entire workflow, from the moment cells are placed in culture. With the SX1, an individual user can improve their cell culture quality control, ensure quality downstream analysis, and generate an information-rich analysis at a scale that is not practical with traditional cell analysis technologies.
I said that it must be very exciting to see all these new uses for the IncuCyte being developed and asked based on what she has seen, if she had thoughts on the future of live cell analysis. Kim said there is so much to look forward to. We are seeing our users embarking on exciting fields like cell therapy and personalized medicine, and taking advantage of game changing technologies like CRISPR. It’s not just about seeing how a cell responds, it’s about manipulating and leveraging all that a cell has to offer. When you start realizing we have everything we need to combat disease, repair an organ, or determine what treatment we need in our own bodies it just fills you with such hope and excitement for the future!
I closed the interview by asking if there was anything else she wanted to add for our listeners? She said that expanding the IncuCyte portfolio is a big step towards fulfilling our vision that every cell biologist can gain live-cell insights with an IncuCyte.
For more information about the IncuCyte portfolio of products, please see essenbio.com/incucyte
Jun 24 2019
Rank #15: Successful Methods for Perfusion Process Optimization
In this podcast, we interviewed Dr. Andreas Castan, Principal Scientist at GE Healthcare Life Sciences about the best methods for optimizing perfusion processes. This included a discussion of tools for media optimization and innovative cell separation techniques.
I began the podcast by asking Dr. Castan what makes perfusion a good manufacturing platform? He identified several reasons including short residence times in the bioreactor through adding fresh media and removal of spent media and as a result is perfect for unstable molecules. Perfusion permits processes to be run with high volumetric productivity by maintaining high cell densities for a long period. With perfusion there are many opportunities for process intensification and it is a good way to quickly get started with production, provided you have a good batch media that cells like.
Next I asked if there were any specific product types or situations that are an especially good fit for perfusion? Andreas described three primary areas – unstable products, integrated continuous manufacturing platforms and process intensifications for removing bottlenecks in manufacturing. Examples would be cell bank manufacturing to create high density cell banks, seed train to remove steps or produce high viable cell density inoculum or hybrid processes of both perfusion and fed batch technologies.
We then discussed the importance of optimization for perfusion processes and the various approaches for optimization. Andreas said it is important to optimize processes to gain the product quality and the process economy that you are aiming for. Cell culture media is the most important thing to optimize in perfusion culture. In order to achieve good throughput for your optimization it needs to be performed in a scale down model then results must be verified under bioreactor conditions.
Then I talked to Andreas about a recent poster he authored and presented at ESACT. The poster outlines development of perfusion specific media and designing medium that supports low cell-specific perfusion rates. I asked him to explain what cell-specific perfusion rate (cspr) is and why a low cell-specific perfusion rate is important? Andreas explained that cell-specific perfusion rate is the volume of media added per cell per day. If you assume your medium supports a cspr of 50 picoliter and you want to run the process at 30 million cells/mL your volumetric perfusion rate would be 1.5 bioreactor volumes per day. If you take the same medium with 100 million cells/mL you would need a volumetric perfusion rate of 5 bioreactor volumes per day, which is not feasible for production. A medium with a better depth of cspr of 10 picoliter at a cell density of 100 million cells/mL would have a volumetric perfusion rate of 1 bioreactor volume per day, which is feasible for production.
His team investigated screening methods to develop medium with a very low cspr. They started with basal media, then screened different feed solutions and Cell Boost feeds in batch mode. The Cell Boosts with a positive impact were taken to a DOE study in spin tubes that replicated perfusion conditions, then promising formulations were moved to perfusion processes in WAVE or the XDR bioreactors.
Next I asked him if you can turn a fed-batch medium into a perfusion medium? He said yes, transferring a fed batch media to a perfusion process is a simple process. He lays out the methodology in the previously mentioned poster and they have done this for two cell lines and three fed-batch media and feeds. The methodology is also described in the Biotechnology Process Journal Article, “Repurposing fed‐batch media and feeds for highly productive CHO perfusion processes.”
I followed up by asking how did the methodology work in the case study presented in the poster? Andreas said it was a very fast method with batch screening taking one week, the DOE study took two weeks and within one month they had the composition for a perfusion media that fit the desired clone. They tested two media developed using this process on their internal Herceptin producing cell line and for both media they reached cspr below 20 picoliter. They were able to reduce one media to only 7 picoliter. The cell specific productivity for this process was comparable to what they saw in fed-batch.
We then discussed another approach to optimizing perfusion processes, optimizing the equipment used in process. In another poster presented at ESACT, Andreas presented the use of hydrocyclones for cell separation. I asked him to describe how hydrocyclones work and why they make an attractive alternative to traditional cell separation devices? He said that hydrocyclone devices are comprised of cylindrical and conical parts that allow centrifugal separation provided by feed suspension introduced tangentially at high flow rates into the device. The absence of rotors or moving parts make it an interesting separation alternative to perfusion in long term operation. A simple device that is not prone to clogging as is usually seen with filtration devices
I then asked what his experiences were with the hydocyclone in the case study? Andreas explained that they investigated separation efficiencies at different flow rates and different flow concentrations and found that separation efficiencies of 70-80% could be achieved with the current hydrocyclone. This test was run using a perfusion process in an XDR 50 bioreactor at 50 million cells/mL for more than 2 weeks in very stable conditions. It was a very successful test of the device.
I summarized by asking if companies are limited in time and/or resources for optimizing their perfusion processes, what would he focus on first? He said media should be focused on first because as explained earlier a low cspr is key to reaching high volumetric productivity, low volumetric perfusion rates and high product concentrations, all of which result in good process economies. Next you could think about how you could use perfusion to reduce process scale.
Lastly, I asked if he had anything else that he would like to add for our listeners. He said that before moving into perfusion you should ask yourself what makes the most sense for your process. You need to think about your infrastructure, prerequisites, platform, and previous knowledge. Then you can decide where perfusion makes the most sense in your process. In cell bank manufacturing, seed train, or production bioreactor. Media is the most important factor to work with and it is not difficult to use existing media and cell boosts to develop perfusion media that fits your clone.
Jul 23 2019
Rank #16: Integrating bioprocessing steps to improve efficiency and reduce cost
In this podcast, we interviewed Dr. Rick Morris, Senior Vice President, R&D, Biotech, Pall Life Sciences, about integrating bioprocessing steps to improve efficiency, reduce cost and address current challenges in the industry. Specifically, we looked at how upstream and downstream can be more integrated and how to increase integration in existing facilities.
Feb 19 2019
Rank #17: Optimizing Fed Batch Culture – Developing New Tools and Methods to Improve Production
We interviewed Dr. Aline Zimmer, Head of R&D, Advanced Cell Culture Technologies at Merck about the importance of optimizing fed batch cell culture for successful biomanufacturing. Aline discussed how her team has developing new tools and methods for optimizing fed batch culture and the impact this has had on productivity and product quality.
May 20 2019
Rank #18: Mustang Bio’s gene therapy has tremendous potential to cure XSCID or Bubble Boy Disease based on study results from St. Jude
In this podcast, we interviewed Dr. Manny Litchman, President and CEO, Mustang Bio and Dr. Knut Niss, Chief Technology Officer, Mustang Bio about the exciting study results for their gene therapy candidate to treat X-SCID, why this disease is a good fit for gene therapy and next steps.
To begin the podcast, I asked Dr. Litchman about the study results in treating X-SCID or “bubble boy disease”, including current standard of care and why gene therapy is a good fit for this disease. He explained that X-SCID is an X linked rare genetic disease with mutation in gene responsible for insufficient immune system generation at birth. As a result, severe, recurrent and opportunistic infections occur and usually results in death by age one if left untreated. Current standard of care is Immune reconstitution by allogeneic stem cell transplant, however the transplant must be from a matched donor and the best donors are siblings. There are only about 15% matched sibling donor transplants at this time. Gene therapy would eliminate the need for a matched donor and would improve survival and quality of life for the patient.
Next I asked Dr. Niss about previous attempts at treating this disease with gene therapy and how their therapy is different. He described that in the early days of gene therapy gamma retro virus was sometimes used as the vehicle for delivery. For the current gene therapy a lentiviral construct is used, which has safety features that the gamma retrovirus didn’t provide. In the previous attempt to use gene therapy for X-SCID, the patients were cured of X-SCID, but developed leukemia due to the use of the gamma retrovirus. Today gene therapies build many safety features into the construct, many of which are incorporated into the vector used in the current study.
I then asked Dr. Litchman to elaborate on the St. Jude study and the very positive results. He explained that the results were published in the New England Journal of Medicine, “Lentiviral Gene Therapy Combined with Low-Dose Busulfan in Infants with SCID-X1,” by St. Jude. The study was a single arm study in newborn patients who were diagnosed with X-SCID. The results were striking as all patients had multi-lineage immune reconstitution. In addition, all patients that had infections prior to gene therapy cleared completely, with levels of IgM that are produced in B cells normalized in 7 patients. Four patients were able to stop IV immunoglobulin (pooled antibodies that these patients receive to prevent infections) and 3 patients had a normal response to childhood vaccinations. The safety profile was excellent with no leukemia and no transfusions required.
Next I asked Dr. Litchman if he could tell us about the two populations receiving the gene therapy currently and other patients who may benefit. Manny said that the therapy is currently being given to newborns with the disease. The incidence of the disease is very low with 1 in 225,000 live births globally. This results in about 20 patients in US. There is a larger population of about 400 who have received hematopoetic stem cell therapy and are eligible now or in the future for gene therapy because their condition has deteriorated due to infections. The National Institutes of Health (NIH) has a current trial running for these patients. Five patients have been treated and they are enrolling more patients and currently have a wait list.
I followed up by asking Manny what are the next steps for this therapy. He said that they will complete the technology transfer of the cell processing from St. Jude to Mustang’s cell processing facility in Massachusetts. It is a state of the art processing center run by Dr. Kniss. They will also continue the ongoing trial and will file for approval. At the same time, they will expand the number of sites sites for the NIH study to a multi center clinical trial.
I then switched gears by asking about their biomanufacturing approach. I began by asking Dr. Niss why they selected a lentiviral approach. Knut clarified that St. Jude selected lentivirus. He explained that it is well recognized that gamma retrovirus not the best approach for stem cell based therapies. He said we understand the integration of the lentivirus into the genome at a much higher level than the gamma retrovirus and gamma retrovirus integrates in a true random pattern where lentivirus has a defined integration pattern. For a therapy like this where you want durable outcomes, it is important to have stable integration into the genome so that the gene is expressed long term.
I then asked what were the biggest successes they found in manufacturing and what their plans are for scaling up in the future. Knut said that they don’t anticipate a problem because it is a fairly simple process. He explained that they isolate CD34 cells from the patient add the lentivirus and prepare the therapy for infusion. Capacity can be built up very quickly. He said that tehy are constantly scouting new technologies looking for the next technology breakthrough. In general, he said that cell and gene therapy products are now not the ultra rare niche technology. Technologies specifically developed for the needs of cell and gene therapy manufacturing have been developed and that has improved cell processing greatly
I asked what is next for Mustang. Manny said that they are looking for other in-licesning opportunities from the academic world of late pre-clinical or early clinical rare diseases that could take advantage of our cell processing facility. In the near term, they have a rich portfolio in CAR T for oncology with 3 therapies for hematological malignancies and 3 for solid tumors. They will be filing 2 INDs this year for two of those programs. One for AML and one for a multiple myeloma target.
I closed the interview by asking if there was anything else that they would like to add for our listeners. Manny said that they were very happy to have completed their capital raise and have at least two years of cash to execute on all their clinical programs. They are also grateful to their collaborators, St. Jude, City of Hope, Fred Hutchinson Cancer Center, and Nationwide Children’s Hospital.
Jul 30 2019
Rank #19: Cell Therapy Manufacturing – Opportunities and Challenges
In this podcast, we interviewed Dr. Robert Preti about the past, present, and future of global cell therapy manufacturing. We also discussed major industry milestones, manufacturing challenges, and opportunities for the further improvements.
View show notes at cellculturedish.com/cell-therapy-manufacturing-opportunities-challenges/
Robert A. Preti, PhDChief Executive Officer and President, Hitachi Chemical Advanced Therapeutics Solutions, LLCGeneral Manager, Regenerative Medicine Business Sector, Hitachi Chemical Co. Ltd.
Dr. Robert “Bob” Preti is the co-founder and the visionary behind the successful growth and development strategy of PCT (on October 2, 2017 renamed Hitachi Chemical Advanced Therapeutics Solutions, LLC or “HCATS”) over much of the last two decades. Upon PCT’s acquisition by Hitachi Chemical Co., Ltd. in May 2017, Bob’s role expanded to General Manager, Regenerative Medicine Business Sector of Hitachi Chemical, responsible for development, management and oversight of the global business operations of Hitachi Chemical’s regenerative medicine business unit. Bob built HCATS to meet a recognized need for high quality manufacturing and development services in an emerging industry. Bob is currently Chairman for the Alliance for Regenerative Medicine, where, among other activities, he previously served as Vice Chairman and in other roles. Bob holds a BS degree in biology from Fordham University, and a MS degree and Doctorate, both in biology, from New York University.
Mar 17 2018
Rank #20: Raw Material Quality – Identifying and Mitigating Variation
In this podcast, we interviewed Cory Card, Principal Scientist Life Sciences, GE Healthcare about raw material quality, its impact on cell culture and technologies to identify and mitigate raw material variation.
Sep 17 2019