Hi everyone and welcome. My name is Katie Silbala and I will serve as your moderator today. Thank you for joining our webinar Make More Lentivirus and Make it Right the First Time. As your moderator, it is my role to ensure that we make the most of your time with us. Before I turn things over to our presenter, I'd like to cover a few housekeeping items. At the bottom of your screen are multiple application widgets you can use. There you can also find our reaction button, indicated by the thumbs up emoji, that allows you to give immediate feedback on the presentation, topic or anything that stands out. All the widgets are resizable and movable, so feel free to move them around to get the most out of your desktop space. You can expand your slide area or maximize it to full stream by clicking on the arrows in the top right corner. If you have any questions during webinar, you can submit them through the Q&A picture. We will try to answer this during the webinar, but if a more detailed answer is needed or if we run out of time it will be answered later via e-mail. Please know we do capture all questions. The webinar is being streamed through your computer so there is no dial in number. For the best audio quality, please make sure your computer, speakers or headset are turned on and the volume is up so you can hear the presenters. An on demand version of the webinar will be available after and can be accessed using the same link that was sent to you earlier. And now I would like to introduce today's presenter. Katish Fair is a Director of Sales and Account Management responsible for developing and guiding strategic partnerships with cell and gene therapy innovators for our viral vector CDMO in Carlsbad, CA, USA. Kathy has been in the biopharma industry for over 15 years, focused on identifying innovative solutions to help biopharma companies bring their therapolitics to market. She holds an Ms. in chemistry from Pruitt University. That's it from my side. It's my pleasure to turn things over to Katy. Thank you so much, Katya. I really appreciate the introduction. And before we delve into our Lantivirus platform, let's highlight some aspects of current Lentivirus production and challenges we are facing as an industry. While I think we can all agree that AAB isn't isn't important for the long term outlook of gene therapy, Lentivirus remains a leading vector segment for the near term as the preferred delivery platform for gene modified cell therapies. It's high volume load, reliable and stable gene delivery ability, and low immunogenicity make it ideal for targeted oncolytic immunological therapies and, increasingly, genetic disorders. That said, the physiological and physiochemical characteristics of lentiviral vectors create some challenges during production. Common concerns throughout the industry we often hear about can be grouped into 3 broad categories. Safety. Performance and purification stability for safety. A lot of times when you think about it, what what are the big questions that haven't been answered? How do we achieve total and functional vector measurement? How what assays can provide virus safety assurance and how do we ensure product and process impurity removal? As an industry, I'm not sure we have solutions to all of those questions, but throughout this presentation you'll hear the solutions we've tried to create that at our company to to help our our customers meet their needs for performance. You know, how can we maximize tighter Current practice is to use a multiple packaging plasmid system, but is there a better way? How can we overcome physiochemical conditions in order to maximize lentivirus recovery and functionality? And from a purification and stability standpoint, how can we reduce high cost of key steps like affinity affinity chromatography? What's the best final formulation buffer that maintains a stable mono disperse population? And how can we overcome thermal instability and freestyle sensitivity? Stick with me and we'll find some answers and solutions that we've created to those questions. For all these questions and challenges, it's important to find those solutions. The cell and gene therapy market is continuing to see significant activity and growth. As you can see by the the charts on this slide. Much of this stems from the growing confidence in the field based on recent FDA and EMA approvals, positive clinical trial results and the potential to provide new curative treatments in the case of lentivirus, especially with oncology being a top focus for gene therapies, there's been a whopping 600% increase in the number of clinical trials over the past five years. There's a strong pre clinical pipeline as well using lentivirus for gene modified cell therapies for cancer indications and a growing number of non oncolytic indications using using lentivirus and all these are continuing on an upward trajectory. The industry is looking for ways to alleviate and rectify some of the challenges we've we've highlighted here and to better support drug product innovators in bringing their therapeutics to market. The focus as of late has been shifting from adherent to suspension cell culture manufacturing and with that the optimization of the transient transfection process to help improve lentivirus production outcomes. The goal here here is to de risk the manufacturing process by providing a scalable defined workflow that provides high yield and recovery, reducing time to manufacturing yields, cost savings and has analytical tools to support the the process. And likewise, we have taken this approach in developing our Virus Express lentivirus prep platform, which I'll detail in later slides. And for the future, over the next few years, we're expecting to see the adoption of technologies that further optimize the process and performance to increase productivity and reduce COGS as well as time to get the therapies to the patients. Some of these solutions that are in infancy are stable production systems, process intensifications, and rapid quality evaluations. How can we help? Well, by leveraging our legacy and foundation built in our deep experience in viral vector development and manufacturing and by developing solutions to overcome these challenges. We have had three decades of expertise in viral vector development and manufacturing. We have experience supporting 4 commercially approved gene therapies and we have been producing viral vector since the gene therapy industry was in its infancy and to date, we have produced over 1000 GMP batches for nearly 300 clients. Our culture is focused on quality compliance and rooted deeply with a responsibility to the products we manufacture as well as to our customers and patients they serve. Our manufacturing facility has completed 9 successful inspections by 6 global regulatory agencies, including the FDA and EMA. Since 2017, we have done a lot of work with Lentivirus for our customers with over 300 GMP batches produced. We've supported within that two BL As and 10IN DS for multiple customer programs. It's this foundation in experience and know how that we draw when we optimize our Lentivirus platform, allowing us to support our our customers with a streamlined and reducible platform. And just to highlight a couple of the the the key aspects of the platform and we consistently see titers in the four E 8 range, which is industry leading. And I'll show you on on in subsequent slides what that means. We're able to provide reproducibly a 20% recovery after final filtration. We've scaled it to 200 liters and beyond. And this process is provides a streamlined path to manufacturing in under 12 months. So it really helps accelerate your therapy to patients as as fast as we can. So how can we best support our customers is always at the forefront of everything we do. Our platform allows our clients to benefit from AD risk approach that provides high quality results to achieve market success. So how do we demonstrate that we have proven performance? I've mentioned before we've this has been used in multiple BLAS&INDS and we've used it for more than five client programs. We're able to accelerate timelines allowing us to start GMP manufacturing in under 12 months. It also provides cost savings because we've been able to optimize the raw materials and the runs are more consistent and and productive. We've also developed a comprehensive analytical toolbox and you can rely on these methods to consistently characterize the product and the productivity, the yield and the purity. And and lastly, you know we have the regulatory support services that provide a customized support at each critical time point and milestone as the therapy progresses through the regulatory pathway towards commercialization. Excuse me, we have developed and optimized our virus Express Lentivirus lentivirus platform with two goals in mind. One is to make more virus and two is to make it right the first time. So how do we achieve that? Let's start with the plasmids. Our packaging plasmids are available off the shelf and available for use and have been optimized to our process. And then the next slide, I'll show you how we've been able to to do that. We have established partnership with an external CDMO to allow us to manage and support the manufacturing of the gene of interest plasmid from our customer. So we take all of that along with our know how. The other elements of the virus Express platform are the HTK 293T lentiviral production cells which are well characterized, GMP banked. It also has a chemically defined media, which is the Excel HTK 293 viral vector media and we take all of that together to make up to develop the platform that delivers consistent results for our customers. We also are able to scale up to 2000, up to 1000 plus liter and we have automated fill finish as part of our process. We have the analytical toolbox I've mentioned as well as regulatory services that really support you from from beginning to end of your product life cycle. So our our platform is designed to support I and D quickly. And because we have optimized the platform from end to end and increased productivity and titers, the total operational spend is lower, which can really help drive down your cost per dose for the patient. So I mentioned I would show you how we've created the plasmids that really have increased productivity. So you can see when we started our our platform approach, we had commercially available plasmids, packaging plasmids and we had an internally developed transfer plasmid. So we've assessed a myriad of of scenarios of commercially available versus internally developed and we found the right combination really increased our performance nearly 20 times over our original platform. So we're very excited to be able to offer our packaging plasmids to our customers to really help get the the productivity results that they need. So not only do we have our plasmids to support you, we, we do manufacturing assessments along with our platform technology to really ensure that we can offer the full spectrum of viral vector development and manufacturing needs. Today we're going to focus on our suspension manufacturing capabilities, but please reach out to us to learn more about our other service offerings and how we can support your preclinical to commercial needs for viral vectors. So using our packaging plasmids in addition to our H EK293T GMP bank cell line, our chemically defined media, our optimized upstream workflows, we were able to achieve best in class performance with a 3X increase over competitors advertised titers. We are so ecstatic to be able to offer that to you. We've used DOE studies to be able to identify the impact of transfection parameters on infectious titer and productivity to identify the ideal levels of plasmids to use during transfection. And through these studies, we were able to achieve titers that have consistently hit the MIDI 8 transducing units per mil, which is is by far head and shoulders above the industry. These identified conditions have been run at larger production scales. So we've not only confirmed it at a small scale, but we have consistently seen reproducible productivity at a larger scale as well. So from from our bench top experiments all the way through clinically relevant production scale, the Lentivirus platform is a robust, scalable, reproducible high titer solution for lentiviral production. And here again, we're just illustrating the MIDI 8T transducing units per mill titers that we've seen. So during the lentivirus platform development, we didn't only focus on the upstream, we've also looked at how we could improve the downstream workflow to maximize recovery. We focused on four key areas and sorry, three key areas, anion exchange chromatography, TFF and sterile filtration unit operations as we've seen that those were identified to have the greatest impact to step recoveries. For anion exchange, we optimize the sodium chloride concentration in the illusion step and determine which condition enabled the highest infection infectious particle recovery. For TFF, we optimize both for concentration and dye filtration and for steroid filtration we looked at the formulation buffer optimization. We looked at a range of solutions to understand the effect of pH, salt stabilizers, hold times and temperatures which all may occur during final processing and storage of the lentiviral vector. So the outcome of that work is seen on this slide. And as you can see, we have high repeatability and consistent step yields as we have been able to achieve overall 20% recovery after final filtration. These identified optimal conditions have been run at larger production scales to confirm scalability and consistency as well. So how is this relevant? You know, we, we talked a lot about how we've improved the, the platform, but what does that really translate to the to the customer and and ultimately to the patient? So we've done some calculations based on the 50 and 200 litre scales that we've performed and we have some assumptions that you can see listed in in the footnote on the on the on the slide. So based on our productivity of the mid E8 transducing units per mil range for your harvest titer we have estimated the number of doses that we think based on an Moi of three what what you would achieve using our platform. So for the 50 liter scale, you know it's it's approximately 1300 and for the 200 liter scale it's approximately 5000. So that's very exciting to conceptualize and to think about South. Again, these are just estimates, but we're trying to give you the visual of what the high tiders were describing could translate for you and the patients in the end. So accelerating to GMP is always a consideration. And when you think about options for taking your program to commercial, you have a bespoke traditional timeline from concept all the way through commercialization or you can use a more templated platform approach like our Virus Expression Express. So we looked for ways that we could gain time savings and optimize the workflow. And this timeline really shows you that from a customer project beginning from process development all the way through GMP manufacturing, it shows you the efficiencies you can gain. So the portion on the bottom that is highlighted as virus express timeline that actually came from one of our our customers. The customer came to us and said I need to have I&D material by Q4, can you help me? And we absolutely could. We were able to begin negotiations in Q1, use our manufacture ability assessment to really speed things along in parallel. And then we were able to do our our PD and prepare for GMP in under 12 months to deliver the GMP product to the client ahead of our schedule as compared to a bespoke timeline that could take as long as three years. So in under 12 months versus closer to three years, it's a huge savings for you all. We also offer a complete analytical toolbox that I've mentioned to support lentivirus characterization and provide the right data about your program and product. We've developed more than 18 methods that are outlined here to ensure that the lentivirus is is safe. It's it's efficacious and it has high quality and that you can trust that it's a robust material for your trial. And depicted on the top is a product life cycle from innovation, you know, from proof of concept all the way through the delivery of the GMP material to the patients we serve. From our manufacturing assessment approach and virus express platform technologies to our regulatory support and fill finish services, we offer the full gamut of viral vector development and manufacturing needs. In the boxes on the lower half are the segments of our industry. Our team of viral vector experts are accomplished in the development and manufacture of a diverse range of viral platforms such as AAV, Lenti, adenovirus and less common vector types like HSV, Rio Echo virus and more. This allows us to support a range of therapeutic approaches from in vivo gene therapies and oncoliticate therapies to gene modified cell therapies as well as gene editing therapies. And regardless of the therapeutic approach and the viruses you utilize for your gene therapy, we have services to help you. Our viral vector manufacturing facility is located in Southern California, about 35 miles north of San Diego in Carlsbad, CA. Our suspension facility is just down the street from our adherent manufacturing facility. And collectively the facilities house process and analytical development, clinical through commercial production, quality control, product storage, all in a single location. Also on site, we have support functions like quality engineering, state-of-the-art warehouse and critical utility systems. And the foundation of this facility is built on our three decades of experience in viral vector development and manufacturing. And no matter where you are in your drug product development cycle, we have the right capability to support your therapeutic. Whether you have a tech transfer process or you need complete development from from the gene and the dream, we are able to support you. And one of the remarkable things about our facility is our on site process and analytical development services. This not only allows us to work on your program from an early stage all the way through to clinical and commercial manufacturing without having to leave our facility in our teams. It also facilitates an interaction and exchange of of knowledge between our development, engineering and manufacturing teams to enable a smooth, integrative and effective tech transfer and scale up for your program. So our process development teams incorporate a customizable virus specific holistic approach in developing that robust, scalable process. We use automation and small scale studies to reduce risk and accelerate timelines as well as help optimize the upstream and downstream workflow that works best for your gene of interest on our excuse me, our on site pilot lab helps to ensure our developed processes and its performance. They're well characterized at multiple single use by reactor scales. You can see that we have multiple, multiple scales and we also have the ability to use those scales in our pilot lab. So yields and product quality profiles achieved in the representative lab and pilot scale are translatable to our large scale production runs because we use the same equipment. And using our pilot lab, we provide assurance that your process is scalable, alleviating that risk of unforeseen scale up challenges, hampering timelines and providing an efficient, streamlined and de risk tech transfer from PD to manufacturing. So our analytical development team supports assay development method validation from analytical through characterization as well as comparability studies to provide accurate and reliable methods for each day's development, which will ultimately transfer easily to manufacturing. We support the analytical team supports process development do ES as well as in depth virus testing with advanced characterization capabilities. Our custom design manufacturing facility was purpose built to help de risk viral vector production and provide suspension cell culture manufacturing through our flexible operations and suite models. Our processes are fully scalable from vial thaw through downstream and fill finish. Our current scale supports 50 liter to 1000 plus liter to cover the life cycle of your molecule from clinical through commercial. The fill finish sweeps feature a state-of-the-art dual chamber isolator and fully automated fill line capable of 3600 vials per hour. A typical 1.2 mil fill volume runs at about 24 miles per mil with 100% weight checks. Our flexible fill line can fill a range of vial sizes from 2 mils on up and our isolator features grade AVHP sanitization, which means FDA and EMA regulatory requirements. Our fill finish area feeds directly into the visual inspection and labeling suite. So it's it's a very nice flow to support the aseptic fill process. I've mentioned a few times that we have a manufacturability assessment and we are we really like to start with this approach as we view it as a key first step to identify and evaluate process improvements. And during the six to eight week assessment, we thoroughly review the process and and assess different areas for improvement as well as perform feasibility studies to test and develop a strategy of using our platform that will deliver the best results for your therapeutic. This process runs in parallel to the proposal process, so it really allows us to be agile and get a jump start on your program. There are many parameters that could be key or critical for transfection optimizations. And a huge advantage of using a DOE approach like we do at our facility is that by taking this approach, you're able to study multiple parameters at a time as well as their potential interactions on each other by using statistical software for experimental design, design and data modeling. This helps minimize the number of experiments that you would be required to perform as compared to A1 parameter at a time approach. We've listed some points to consider here on this slide when we develop our DOES for experimentation. So as part of our transfection process and improving our platform, we explored transfection VCD, transfection reagent to DNA ratio as well as total DNA concentration. And using this set of experiments, we used the standard PEI based transfection reagent and we used a design, a type of design called response surface, which allowed us to generate a model to determine the optimal condition for our transfection parameters. The graphic on the left is an example of the prediction profiler from jump, which shows how the process or how the responses change as the levels of the factors change. So using the model, we were able to identify a condition that resulted in approximately a 60% increase to our genome titers compared to our starting baseline condition, which was extremely exciting and very gratifying for those folks that were invested in this endeavor. So our dedicated viral vector experts are here to help you and bring your novel treatment options and potential cures to patients. Working with a trusted CEO DMO partner can help your cell and gene therapy reach patients more quickly. And when working with us, you can leverage our preclinical to commercial solutions, our strong regulatory track record and quality systems, our established processes and our scale and capacity to accelerate, accelerate your therapeutic to market. The Virus Express platform for Lenti viral vector leverages experience, knowledge and expertise from almost 3 decades of viral vector development and manufacturing to provide a de risk reproducible and high quality path to clinical and commercial manufacturing. This fully optimized approach has already been used to accelerate production and seamlessly scale up lentiviral based gene therapy programs through Ind submission into clinical trials and leveraging our deep viral vector expertise, we have optimized and divide and defined our upstream and downstream processes to deliver high tiders per batch as compared to competitors. We've been able to conserve time and resources for our customers. We can help deliver lower cost per dose therapies. We've been able to accelerate timelines while reducing risk. We have the quality to achieve market success. We've been able to streamline and and make our workflows scalable. We have comprehensive analytics to support you as well as regulatory prowess to really see your process through commercialization. So with that, I will turn it back over to Katya, I believe. Thank you, Katy for this great presentation. Now it's time to answer a few questions that have come in from our audience. But before we do, I would like to remind you that it is not too late to send us your questions now using the Q&A widget. We will try to get through all of them, but if we run out of time, we will respond to you individually. As a reminder, this webinar will be available on our website soon. All participants will receive an e-mail notification when it is available for viewing. Now back to our speaker who will start answering questions that have come in. And here is their first question from our audience AT where are the plasmids coming from? So that's a great question. We have a Bank of packaging plasmids and those constructs were developed internally and we have a partnership with a leading high quality plasmid DN, A/C DMO and through that partnership we manufactured a Bank of GMP plasmids, so they're available to use. Thank you so much. I see question already. What nuclease do we do you use and what are the salt conditions in which it operates? The the nuclease that we use is Benzonase and typically we have found that if we do a single step pollution between one and 1.5 molar sodium chloride we we see improved recovery and we have high coastal protein removal as well. One more interesting question from our audience, what is the overall quality for a Virus Express lantivirus platform? That's a another great question. Our platform has been able to achieve consistently 20% recovery after the final filtration. So we are able to consistently produce high quality and and higher yielding batches. Because of. Because of our downstream capabilities. And can you test for Z deals? Absolutely. We we've been able to develop some methods and and establish them. We're able to test not only for raw materials, but also for the cellular debris. So we have methods to test for residual benzonase as well as residual host cell protein post cell DNA. So we can we can understand the impurity profile very well. There is one more question just just came in. What is the status of stable cell lines in general? That's a a really interesting question. I know there in the market there are a few stable producing cell lines, some are specific for Lentivirus, some are specific for getting AAV. At at this time for our capabilities, we are assessing and and you know looking at several different options to see what what is the best fit for our capabilities. But at this time it's it's not our in our current offer. Thank you so much. And what is the ideal phase to start a manufacturing assessment with our platform? So for the manufacturing, for the manufacturing assessment, really any phase is ideal to begin, but to make the most time to to make the most of your time to market and to establish the best process early on. Best time is to start at the very beginning when you're developing your construct early on. This will save you time and money as your therapeutic moves through towards commercialization. So that can really help you. The manufacture ability assessment can really help determine the best. Process to to. Support your program and meet its milestones to reach those patients after commercialization. And what if you are a maximum field size? So our equipment can handle multiple different fill configurations and sizes. So we don't really have a limit. We can handle multiple different kinds of fill containers. You know, if you have a vial or if you have a, a bag, we could certainly and the correct configuration to fill your container are really our, our limiting factor is how stable is the virus and and how long will it take us to visually inspect the material so that we can meet pretty much any requirement you have as long as it's stable enough and inspection done in time. There is one more question that just came in. Do we need to pay royalty if we use your plasmids? That is a great question and no are our plasmids are royalty free. And what is the status of stable cell lines in general? I think, I think we already answered that question. So there are a few stable cell lines in the market both on the lentivirus side as well as on the AAV. From from our capability perspective, we are currently assessing several that that we are considering if they're a good fit for our our capabilities. So stay tuned. We'll. We'll have an answer for you on that hopefully by the end of the year. Thank you very much for all these questions. If we didn't get to your question, please feel free to e-mail our presenter directly the register For future webinars or to access our aircraft webinar library, please visit our website. You can also click on the calendar widget at the bottom of the screen to register for our next webinar. I would like to thank Katie for today's presentation and thank you to our audience for joining us. Have a great day. _1732109223284

  

    

_1732109223547