Navigating Industry Challenges to Drive Sustainable Plasmid DNA Strategies

Arsalan Arif (00:02):
Hi, everyone. Arsalan Arif here with Endpoints News, and thanks for joining us today for navigating industry challenges to drive a sustainable plasmid BNA strategies. We're sponsored by Charles River, and I'm excited to moderate today's discussion. Joining us today we have Andrew Frazer, the Associate Director of Scientific Solutions at Charles River Laboratories. Now if you have any questions during today's webinar, be sure to hit the Q&A button at the bottom of your screen. We have reserved some time today to get to your questions. This session will be available on demand tomorrow to rewatch or to share it with your colleagues. Now, I'm going to pass it off to my guest, Andrew Frazer to get us started. Andrew?

Andrew Frazer (00:43):
Thanks. So, hello everyone and welcome to this webcast. I'm going to talk today about some of the current industry challenges around plasmid DNA, and what we can do both as suppliers but also consumers of plasmid products, to help drive sustainable strategies. It truly is a very exciting time at Charles River. I'm really looking forward to discussing some of the common themes within the plasmid, but also the wider advanced therapy space, and importantly, some of the developments that Charles River are making as a company recently, in response to these challenges.

(01:30):
I expect many of you will be familiar with Charles River. The company has a very long history and really a worldwide presence with over a 110 facilities supporting capability at basically every stage of drug development. To add a little bit of context, Charles River was directly involved with the development of 86% of novel therapeutics approved by the FDA in 2021. This included around 1300 INDs and IPDs with about 30% of those being within the cell and gene therapy space.

(02:08):
While Charles River is most well known for its history and its capability and testing, the company has established a global network of facilities that span each of the major CDML platforms for advanced therapies, and this includes cell therapy on our Baltimore and Memphis sites. Viral vector at Rockville. And the one that we're going to talk about in more detail today is plasmid DNA, located in the UK at our Alderley Park and Keele facilities. This network, in combination with our testing sites, does provide very much an end to end portfolio of services for advanced drug modalities going all the way from discovery and nonclinical development through to GMP manufacturing.

(02:56):
I hope that brief introduction helps set the scene for Charles River, and really just highlight our vision within the advanced therapy space. But for the remainder of the presentation, I'm going to focus very much on plasmid DNA, and the critical role that it plays in supporting development and delivery of advanced therapies. Some of the common challenges associated with sourcing and supply of plasmid DNA, and how Charles River has responded to these challenges by developing our platform production approach, introducing purpose-built facilities, and also expanding our service offering to include off the shelf plasmid products.

(03:49):
So, when we look briefly at the wider cell and gene therapy market, it's very clear that it's still growing and evolving very quickly. And in fact, one in five new drug candidates fall within the cell and gene therapy category. As more and more products are successfully delivered to market, the growth is extensive and there are huge levels of investment and a growing number of companies involved. The vast majority of which fall within the small to medium sized biotech mainly... And the general or the focus is on genetically modified cell therapies. Some key emerging trends that we can also see include the use of stem cells, and gene therapy like drug candidates like Messenger RNA, with companies getting creative on their business models and also their manufacturing approaches.

(04:51):
While the future looks incredibly bright, the industry is not without its challenges. This slide shows some important information taken from nature reviews and drug discovery, and it highlights the unique position we find ourselves in within the cell and gene therapy space. So, unlike classical monoclonal antibodies, lots of advanced therapies are having issues and complications with CMC. And they experience a much higher rate of disruption for regulatory approval, which ultimately drives up cost and time. When we look at the numbers in a little bit more detail, we can see that many of the issues that we can see or the disruptive issues that we can see stem from manufacturing and also comparability between clinical and commercial supply. And this points to the requirement for stronger validation or early on within programs, but also continuity when progressing from development through to clinical trials.

(06:04):
If we keep this in mind when we look at the role of Plasmid DNA, it highlights the importance of getting it right from the start with your Plasmid supply. plasmid DNA acts not just as a drug product in itself in the case of DNA therapies and vaccines, but also as a critical starting material for delivery of multiple advanced modality therapeutics. If we consider Plasmid DNA in this critical role, or even view it as the foundation for a successful advanced therapies journey, therein lies a challenge but also an opportunity. I've selected some... Or today, for this presentation, I've selected some of the common challenges that we often encounter directly as a manufacturer of Plasmid DNA, and also through feedback and experience working with a wide range of drug developers.

(07:03):
I've then highlighted what Charles River have done as a direct response to help address these challenges, and I'll talk through some of the choices and considerations that developers should be aware of. Although many aspects of Plasmid processing are specific to the type of construct an onward application, it is very rare that we have the luxury of time or budget to undertake extensive process development or optimization. I'll talk a little bit about how Charles River has developed a platform approach to processing and standardized manufacturing and response to these types of common project challenges and all also our client requirements.

(07:44):
Secondly, manufacturers and drug developers need to carefully manage and adapt to an evolving set of quality standards and manufacturing approaches for Plasmid DNA. This one is very heavily linked to our phase appropriate plasmid service offering at Charles River, and also the introduction of purpose-built facilities and accompanying quality systems. The last one that I'll run through is one of the newest developments on our plasmid service offering, and these are pre-made off the shelf plasmid products and how they can address some of the challenges and project costs and timelines, but also some of the secondary benefits that can be realized when we adopt this type of approach to supply.

(08:36):
So, plasmid platform processing and standardized manufacturing. When we look at plasmid DNA production platforms, there is of course variation with manufacturers. They generally all use different equipment. They operate at different scales of production, and quite often utilize specialists in-house technology and know-how. However, they do follow a general framework. In the CDMO world manufacturers generally need to accommodate multiple products, so multiple different plasmids for different applications and often multiple host cell lines for production.

(09:23):
So, processes are often set up in such a way that they can sustain productivity and quality attributes, but limits optimization and deliver quick turnaround and contain overall project costs.

(09:40):
When I began working in plasmid contract manufacturing, it seemed that the majority, excuse me, of client requests that we received included a dedicated period of plasmid specific also in-depth process development and optimization. Generally, we adopted a traditional approach, which incorporated screening, various host cell lines, media compositions. Usually this would've been done in shake flasks. We carried out identity and stability checks, so the most favorable ones, and then progressed through a second stage of development, which focused on driving up product yield through upstream optimization, and then aligning this with product purity improvements through DSP Development.

(10:32):
Once we had established an optimal plasmid cell line and set of process conditions, we would then go into a exercise where we did a process definition and performed a scalability assessment before initiating GMP manufacture. So, this approach was great in that it delivered results, but there's no way to get away from the fact that it takes a long time, and often incurs high costs.

(11:09):
One of the key developments that I guess led to the beginnings of a more standardized platform approach was the introduction of high throughput technology. So, a number of years ago at our Keele site in the UK, we made the decision to invest in systems like the Ambr 250 modular bioreactor system from Sartorius. And this was directly in response to the growing expectation from our customers, that development timelines and costs should be significantly reduced. So in a very short period of time with systems like the Amber and another high throughput analytics, we were able to significantly cut down this type of development package by about 70% in time and up to 60% in cost versus the same scope of work performed in more traditional autoclaveable glass fermentation systems.

(12:02):
They added benefit to adopting systems like the amber was. In fact, the greatly improved data capture, so much higher numbers of experimental replicates. On top of that real-time analysis of in-process data, and importantly the ability to implement quality by design methodologies to determine things like critical process parameters and also determine their impact on product quality attributes.

(12:33):
The position in which we find ourselves today utilizes our track record in working with a wide range of plasmid types and basically it allows us to implement a much more standardized and rapids screening approach for project initiation with the plasmids.

(12:59):
Depending on the plasmid type. Our typical approach now involves a rapid clone screen. This is followed by an optional amber evaluation for specific plasmid types that, for example, require productivity uplifts. And that would identify the most favorable set of pre-established process parameters that can help improve productivity, but also maintain product quality. So this basically allows perhaps not a fully optimized approach, but instead a much more fast track progression to manufacture with very predictable and reliable yields, and a retention of final product quality.

(13:49):
So with this capability at our disposable, we recently launched our eXpDNA, plasmid platform service offering, delivering industry leading timelines, but with very much a focus on process and products quality attributes.

(14:11):
So this slide is, I've included this one because we very much view the foundation of any plasmid manufacturing campaign, or the most important part of manufacturing campaign being the cell bank. And while cell banking is well established from a process testing and also a regulatory standpoint. It is an area that can cause program delay and frustration if it's not managed correctly. While our plasmid manufacturing process is largely standardized and it'll talk about that in the next slide. We have developed an effective set of tools that allow us to efficiently screen a range of different plasmid types that are common to cell and gene therapy applications, and allow us to navigate many of the common issues associated with the generation of reliable plasmid containing cell banks.

(15:11):
So, the first step in initiating a plasmid manufacturing project involves an assessment of the plasmid sequence, and importantly, identification of key features that are included to deliver its function within its intended application. There are many plasmid types that actually would not require any of the steps that are shown on this slide. However, plasmid constructs containing regions that are prone to issues, so things like mutation, deletion, recombination, when we think about those things, we're referencing plasmids like ITR containing plasmids for AV manufacturer or poly (A) sequences that are common in plasmids used as templates for Messenger RNAs therapies. They typically involve some extra steps in order to mitigate the risk of failure at the point of final product testing and release.

(16:11):
So, I've already mentioned how we can utilize the Ambr 250 system as an early stage screening and pre-production tool, to provide an early indication on process performance, also productivity assessment and yield prediction. And where we're required, this type of assessment can now be performed in parallel to other pre-banking and screening steps that involve essentially selection and pre-banking of a number of clones followed by checks on plasmid integrity using next generation sequencing.

(16:50):
It is a system that we've had in place for a number of years now, and actually when it's implemented in full, it has a 100% track record on successful delivery for challenging plasmid types. So, with supporting in-house testing from our wider Charles River network, we're actually now in a position where we can recommend and implement this approach for our customer programs with very minimal impact on the overall project cost and timelines. And importantly, it provides a high level of security for onward processing to compliment, I guess the robustness of our sale banking approach. Our plasmid production process also follows the same principles, so the focus is on rapid delivery and very much a focus on retaining end product quality, and high levels of quality control. And this ultimately delivers a reliable and right first time delivery of purified plasmid.

(17:56):
Our process is not plasmid specific, but instead it has been designed in such a way that it can accommodate a range of plasmid types while delivering productivity that meets our customer's requirements, and also ensures reliability on end products final specifications. I'll freely admit, there is potential to drive improved productivity and higher batch yields, and this generally becomes more important when moving towards commercial manufacturer. We have actually found that the vast majority of our customers place much more value on rapid and reliable production, as well as high quality specifications and also little risk of having to repeat or remanufacture due to failure.

(18:51):
In addition to this production strategy, the standardized nature of our processes has allowed us to simplify our supply chain, so we can now hold significant stocks of long lead consumables, and operate both of our main production processes for HQ and GMP with a fully standardized and single use process stream. So, this includes things like dedicated single use, pre-packed chromatography columns. This carries a number of benefits when we are manufacturing multiple plasmid products within the same facility. So firstly, we negate the requirement for costly, and oftentimes consuming Interbatch equipment cleaning and residuals testing. So, this allows us to more quickly turn around our production suites and make them more efficient. And secondly, the fully single use process combined with our longstanding and tried and tested cleaning and segregation procedures, effectively rules out the possibility of product cross-contamination.

(20:09):
And again, this is something that we recognize as being of critical importance to us as a manufacturer, but also to our customers as it can be difficult to detect immediately for some plasmid types. And it is an area where we commonly receive feedback from customers who've experienced issues with other suppliers who perhaps don't implement the same level of control and segregation.

(20:39):
So, just to round off this section, the slides summarizes our main plasmid service offerings. So we have our research grade plasmid. This one is mainly used for in vitro research and development or preclinical studies. It's produced in an R and D production lab, and it comes with a streamlined panel of analytics to reduce time and cost.

(21:03):
The next step up from this, which is quite a significant step up, is our high quality HQ plasmid. We do have a range of clients who go directly for this quality at a very early stage, but generally it's used for toxicology studies, also GMP vector production for phase one and two clinical trials. And as a template for GMP Messenger RNA. It has a much more comprehensive documentation package, and it's produced in dedicated segregated HQ production suites, utilizing GMP principles. So, that's something that I'll talk about in a lot more detail in the next section.

(21:45):
Lastly, but certainly not least is our gold standard GMP grade plasmid that can be used for all stages of clinical supply. Also, commercial viral vector manufacturing and plasmid DNA vaccines. Our GMP products are manufactured in fully licensed GMP facilities with full documentation, analytics and also QP release.

(22:15):
So, I think it's fair to say that overall this is a very similar service offering to many other plasmid providers. However, again, based on direct customer feedback and experience working with a long list of drug developers, we have adapted our offering to essentially prioritize speed, but also retain high levels of quality control. And importantly, by implementing aligned process unit operations, materials, testing methodologies, we can deliver a more seamless transition between quality grids to support all stages of clinical development.

(23:02):
So, the next item that I'd like to talk about is the utilization of purpose-built facilities and quality systems for plasmid DNA. And this is a subject that is dear to my heart, having worked in the early stages of our Alderley Park, project and now seeing it come to life with our grand opening last month. It's very exciting development within our plasmid service offering.

(23:34):
So, before we get into Alderley Park and what it can do, I thought it'd be good to first remind ourselves a little bit about the history of plasmid DNA regulatory guidance for advanced therapies, and highlight some of the challenges that we've encountered both as manufacturers of plasmids, but also end users in the development of drug products.

(23:56):
In 2005, EMA issued guidelines for the production of lentiviral vectors, and within this, there was a reference to plasmid being high quality. Unfortunately, the definition of high quality was not so clear, and manufacturers were left to determine this for themselves. And there was a range of different high quality grades offerings launched, basically with manufacturers providing their own interpretation of what high quality was. And it left developers in the position where they then had to decide whether this was right for them and then justify the use of these types of plasmids with regulators on a case by case basis.

(24:46):
A key update was in 2007, this when the FDA provided specs for plasmids that would be used for direct human use. For example, in DNA vaccines, the 80% covalently closed circular specification has actually been retained as, I guess, the minimum industry standard for all plasmid DNA products, but many manufacturers might produce plasmids with supercoil levels at much higher specs. In 2018, the EMA determined plasmid as a starting material rather than raw material, and added further guidance around ID Testing. Particular point to note was sequence of the therapeutic gene of interest, and this was actually then taken further by the FDA in 2020 where additional guidance was provided for manufacturing and testing of plasmids used for gene therapy.

(25:44):
So at this point, high quality grade plasmids were, although there were some guidance around it, specifications were developing, it was still a gray area within regulations. And with this gradually evolving set of guidelines, it did make it difficult for manufacturers and also consumers of plasmids to determine clarity on quality standards for their specific application. And importantly, and I guess one of the main concerns was that there was an underlying risk that potential regulatory decisions regarding HQ grade plasmids could change. And this could have an immediate resulting impact on ongoing drug development and also potentially impact the demand for shipping grades of plasmids.

(26:40):
In 2021, the EMA issued a guidance document, which I've just added a little snapshot in the top right, basically reviewing principles of GMP and where these apply in the manufacturing of advanced therapies. So for the first time, there was clear regulatory guidance on the quality standards for plasmid DNA used for all of the major advanced therapies. So this guidance, when you look into it does present a very logical approach. And to be honest, it is an approach that many manufacturers were already following, including ourselves, where the end product be that plasmid, viral vector or modified cells. As it moves closer to the patient, the level of quality increases.

(27:32):
And really, we could spend a long time looking into the details here, but the main takeaway is that GMP grade plasmid is not required when used as a starting material for advanced therapy products. However, for certain materials, it is mandatory that the principles of GMP are complied with. Principles of GMP in itself essentially means that manufacturers of advanced therapies should take it upon themselves to verify that appropriate GMP requirements are implemented for the manufacturing and testing of starting materials using a risk-based approach.

(28:16):
So with this, so I guess as a manufacturer, we now had a clear opportunity to align our HQ plasmid service offering with this guidance. And at the same time, we also took the decision to address some of the new customer requirements that came to light since the initial introduction of our HQ service offering.

(28:38):
So, our plan was relatively simple. We introduced a dedicated HQ quality system adopting principles of GMP. We made it very easy for our customers to adopt either HQ grade production cell banks or GMP Grade master cell banks, depending on their specific requirements. But also importantly, without heavily impacting overall project timelines, we developed and implemented a process that was completely free from materials and reagents of animal origin, which is also something that we've now also aligned within our GMP production process. And as mentioned previously, our manufacturing process stream is fully single use to address the risk of cross-contamination and deliver rapid suite turnaround. We introduced the option to have fully aligned testing for our HQ and GMP plasmid with the option to also include sterility testing and micro plasmid testing, to support onward vector manufacturing activities.

(29:50):
And finally, we totally recognized the need for rapid delivery. So, through some of the things that I've previously mentioned, like standardization of materials, documentation and testing, we've been able to deliver what we feel are best in class timelines for custom plaid manufacturing at HQ grid. And this will be further enhanced by the introduction of pre-made off the shelf products, which I'll talk about in the next section.

(30:24):
Excuse me. So, the culmination of this work was the launch of our dedicated plasmids DNA Centers of Excellence in the UK. Our Keele site has been producing both GMP and non-GMP plasmid products for over 20 years, and it now acts as our dedicated GMP plasmid manufacturing site and is actually currently undergoing its own capacity expansion. Alderley Park is Charles River's newest CDMO manufacturing site. And as I mentioned previously, it just opened last month. The site has three fully segregated and independent process streams. They can all operate in parallel, and this allows us to deliver an excess of 120 batches of HQ plasmid per year with process facility and product quality specifications aligned to the principal's GMP.

(31:28):
The purpose-built nature of the facility allows us to operate under the specific requirements necessary for our HQ service offering. And it importantly allows us to realize a range of operational efficiencies that we can pass on to our customers, versus performing HQ manufacturing in a full GMP facility. It's now operational and audit ready. And really it represents a significant expansion in capacity and capability as Charles River continues to respond to our customer feedback and create a really strong integrated global network to support delivery of advanced therapies.

(32:19):
The last item then I'd like to talk about is again, a very new addition to our service offering. And it's been introduced to further address some of the challenges that our customers face on costs and timelines for plasmid supply, and that's off the shelf products.

(32:41):
So, when we look at two of the main uses of plasmid DNA for viral vector manufacturing, have shown in this slide, we can see that gene of interest plasmids are very much therapy specific and they require custom manufacturer. However, there are other plasmids that are used routinely for different viral vector products. So in the case of AAV, Helper plasmids are relatively universal. And to a certain extent, so are Rep Cap plasmids. However, a range of viral serotypes and combinations of serotypes are often utilized for Rep Cap plasmids depending on application. So, there is potentially a limitation there. Although for Lentiviral Vectors, three of the four plasmids VSV-G, Gag-Pol and Rev, are commonly used with different gene of interest plasmids for different therapies.

(33:41):
So, when we consider this, there is clearly an opportunity to standardize the use of these common plasmids, and manufacture them in bulk amounts made available immediately off the shelf. The aim here is that this will help address capacity constraints that are common in the industry. In turn, that will impact improved lead times that result... Or sorry, restriction on lead times that results from having to perform multiple repeat custom manufacturers. Prices can potentially therefore be reduced. And importantly, customers will have the opportunity to plan and place orders for these common plasmids on a gram versus batch basis, and avoid issues with both under and over ordering and the associated wastage.

(34:39):
If we take a look, a very simple example of a manufacturing campaign to supply plasmids for onward AAV manufacturing, we can clearly see the immediate benefit of off the shelf helper and potentially Rep Cap plasmids, versus fully custom manufacturing on the time and the manufacturing capacity and effort that's required. When we think about this a little bit further, there's also a range of secondary benefits that we can realize.

(35:12):
So certainly, the reduction in complexity of the supply chain, and the ability to reorder common plasmids like Helper plasmids on short notice really shouldn't be underestimated. And in addition, the use of these standardized plasmids does have the potential to ease regulatory approval in the future, and also the introduction of developers follow on pipeline viral vector products in the longer term.

(35:49):
This slide just summarizes Charles River's plan portfolio of off the shelf plasmid products, that they're all set to launch early next year. With our helper and Rep Cap variants that have been used for a number of years routinely for suspension based AAV manufacturer, our viral vector sites. A range of different production scales, so going from 10 to 50, to 200 liter, and also our LVV lentiviral vector, rev gag pole, and VSVG plasmids. So all these plasmids will be available to our customers with full analytical packages, both in HQ and GMP grades for onward manufacturing. Either at viral vector and cell therapy sites in the US, or for your own in-house development or manufacturing requirements.

(36:41):
Looking further down the line, we will aim to develop the service offering to include antibiotic free versions based on our in-house Operator Repressor Titration technology, and also in recognition of the ever-increasing interest in Messenger RNA based therapies. We have began to explore options for inclusion of generic backbone plasmids for linearization.

(37:11):
So finally to summarize, first of all, I hope you find this useful. The intention with this webcast was to get both suppliers and also consumers of plasmid products thinking about some of the common challenges we face, and how we can improve the delivery of plasmid products, and ultimately the delivery of medicines to patients.

(37:39):
In summary, this slide covers some of the common steps that are undertaken when sourcing plasmid DNA. And I've purposefully shown it as a winding pathway rather than a straight line. We can see that the challenge is clearly complex, but we can also see that as the field develops, more and more opportunities are available to help standardize manufacturing streamline things like testing, and supply chain, and the associated documentation to help accelerate some of these wider advanced therapy programs.

(38:14):
I would encourage all developers to think well ahead, plan a strategy that meets both your requirements for initial supply. Very much think about laying a solid foundation through reliable cell banking. Consider the initial quality standard and how that can impact ease of transition when ultimately demand for plasmid ramps up. I think there is no one size fits all approach, but certainly by planning in advance, there are options available that can help minimize later stage disruption that we talked about at the beginning of the presentation, and avoid the requirement for time-consuming and costly comparability studies, and ultimately deliver successful navigation through regulatory approval.

(39:22):
We do very much feel that Charles River's eXpDNA platform provides a range of solutions for common plasmid supply challenges. And when we combine this with our capability in cell and gene therapy and biologics testing, we do have the ability to deliver an integrated solution for the delivery of advanced therapies. And this does include all aspects, so scientific, regulatory, technical and logistics all under one roof. So thank you everyone for listening in. Again, I hope you all find that useful, and happy to run through some questions.

Arsalan Arif (40:08):
Thank you very much, Andrew. Very good presentation for us. And I do have some questions and we have some questions coming in from the audience. I want to encourage everyone in the audience to get some questions in here or Andrew for the end of our session over here. But let me kick it off with some questions from the audience here, Andrew. This question is from the audience here. "Is the NGS conducted in-house or outsourced to a vendor that specializes in that service, Andrew?"

Andrew Frazer (40:36):
Yeah, it's a good question. So yeah, now being part of Charles River, we as a manufacturer do have the opportunity to kind of bring this testing service effectively in-house. And I do agree, it is very specialized, but we do have the ability to perform NGS sequencing in-house. It's not performed on site at our plasmid manufacturing locations. But effectively with having the capability within Charles River, it allows us to carefully manage scheduling of testing, as well as prioritize turnaround for priority samples, and all those things which help us deliver products through release and to customers more effectively.

Arsalan Arif (41:31):
Okay. Here's a question for you. Is serum free media important for plasmid manufacturing?

Andrew Frazer (41:44):
So for plasmid manufacturing, certainly we wouldn't use serum free media. In my experience, that would be something that would be implemented for mammalian cell culture. Although in the same vein as using serum derive products, I did mention in the presentation that there is a customer expectation that we use not on animal derived products. So, definitely that is something that we have implemented both within our HQ plasmid process and also now our GMP plasmid process.

Arsalan Arif (42:21):
Okay. Let me ask you a question here. Do you see high quality DNA being a long-term solution for use in advanced therapy applications? And second part, when do you recommend starting with use of GMP plasmid DNA instead of high quality plasmid DNA?

Andrew Frazer (42:41):
Yeah, it's an interesting question, and it can be a tricky one to answer. I think both of those things are very much related and dependent upon the application. So really on one hand you've got, you can see that manufacturers are tightening up on quality control, and in particular testing requirements for HQ or equivalent intermediate grid plasmid. So, it would be quite easy to say that this trend might continue, and eventually HQ would be phased out and replaced completely by GMP plasmid. On the other hand, we are receiving much better guidance on how to use and utilize HQ grade plasmid for various advanced therapies. And it has obviously provided justification for our developers as well, particularly in the field of cell therapy to push much further forward in clinic with the use of intermediate grade plasmid. And actually now we have directly encounter developers who are planning to take HQ grids right through to support commercial manufacture.

Arsalan Arif (43:51):
Okay. The question here is what technique do you use for super coiled analysis? Can you share an insight there?

Andrew Frazer (44:02):
Yeah, definitely. So we currently use capillary electrophoresis for all of our super cold analysis. Other common assays in the industry are AGE analysis, so AG dens telemetry. It is something that we have previously used for our lower grade plasmids, but as I mentioned before, the new platform is very much focused on continuity, ease of transition. We did take the conscious effort to align testing across those grades, so all of our super coiled analysis and I performed by capillary electrophoresis. Obviously, that's a more sensitive method. It also has the advantage of being able to identify different DN AIS forms as well.

Arsalan Arif (44:52):
Okay, very good. Well, for the audience, keep the questions coming. I've got a couple more here as well from them. Here's the question, I'm going to read it as is, but I might ask you for me. So, "When should developers engage with CDMOs regarding their plasmid DNA needs?" I'm going to guess this as early as possible, or maybe you could just, when is it too late?

Andrew Frazer (45:15):
Yeah, so obviously as a CDMO, we would recommend the developers engage from the very earliest possible stage. That being said, we also recognize that there are many contributing factors that influence the ability of people looking to source plasmid to go to a CDMO. I would say that my recommendation would be, at the very least, go out and speak to potential suppliers. Try to understand what is on offer, and weigh up the pros and cons.

(45:54):
By all means, ask yourself, how far can you potentially progress with your chosen plasmid source? And if and when a change is required, are you actually receiving the necessary information and data from your current supplier to allow an easy transition or movement to another source? Typically, these are the sorts of challenges or issues that cause big delays in clinical programs or development of these advanced therapies, and it shouldn't really be underestimated. So, I think the further you can plan ahead, more information you can get at an early stage, the easier life will be later on.

Arsalan Arif (46:41):
Got you. I've got a couple different questions here related to speed and time. I'm going to ask both of them and then I'm going to ask you to react to them together. So one question one is, "What have you done to reduce the time from proposal to project initiation?" And question number two is, "What has the most impact on timelines, speed and yield, and upstream or downstream processing?"

Andrew Frazer (47:12):
Let me answer the first question first. So this was about proposal to project. So I think it is a really good question, and I'm kind of glad it's come up because it has been a key focus for us as part of this amped service offering on plasmid, because having a background in project management for us Bill is one area where projects can incur significant delays in those very early stages. So the information transfer is incredibly important that that is done as efficiently as possible.

(47:53):
So, when we think about it as a manufacturer, we've got a number of compliance requirements that we need to satisfy to be able to onboard a starting plasmid from a customer. And if these aren't made clear very early on, it does have the potential to be a surprise to customers, and it can cause major delays in our ability to initiate manufacturing activities.

(48:22):
On top of that, I think we found generally that the earlier we can start sharing information on plasmid products, the better as we can start to populate things like risk assessments, even our batch documentation manufacturing plans at a very early stage. And so, we think about all that. We have developed a revised approach to our project initiation. We've done this through feedback from our customers, but also working with an internal cross-functional team of representatives. So, encompassing everything from operations through quality, project management to commercial, and make this a much more streamlined approach and really just to avoid surprises and help avoid those early stage delays. The second question then was around what has the biggest impact on speed?

Arsalan Arif (49:21):
Yeah. Is it upstream or downstream processing? Yeah, speed and yields.

Andrew Frazer (49:26):
Speed and yields. So I think one thing that I would say is that having this standardized platform approach, it has allowed us to clearly predict requirements for projects. And again, it's all about that forward planning. So when we consider upstream yield, it's great to get lots of material out, it's great to have a very good cost of goods. But what's more important than that is predictability. And again, not having any surprises when it comes to delivery of product to customers. So I'd say that those are the things that in my view, would have the biggest impact. I think as long as the yield is good enough, that's what we need to be aiming for.

(50:26):
With regards to the DSP process, obviously this has the potential to impact things like our end product quality, which is actually even more important than anything else. So we absolutely must hit our target specifications as a manufacturer. If not, you're into all sorts of fun games with how specification results, repeat manufacturer, all this kind of stuff. It's devastating for us as a manufacturer. It's also devastating to our customers who are trying to coordinate supplier around clinical programs

Arsalan Arif (51:01):
That's got to be done. Right. All right. We're almost running out of time, but I want to try to get at least a couple more questions in for you, Andrew. So, my next question for you is, when will off-the-shelf plasmids be available and are there any royalties around them?

Andrew Frazer (51:17):
Yeah, so our off-the-Shelf plasmids will be available early next year. They'll initially start off with our AAV helper plasmids, and then the lengthy viral vector package in plasmids. All of these plasmids will be royalty free. So, what I would encourage people to do is keep an eye out in the coming months for press releases. We'll also be issuing technical white paper, showing performance in vector manufacturing.

Arsalan Arif (51:46):
Great. And then, I think my final question for you today, Andrew, is do you also produce viral vectors?

Andrew Frazer (51:55):
Yeah. So as we, I guess briefly touched on just on the introductory slides, we do within our CDMO network have viral vector manufacturing capacity. So in addition to the establishment of our DNA Centers of Excellence in the UK, we also made the strategic decision to focus viral vector manufacturing at our Rockville site in the US. And I'm very happy to say that since we've done that, we currently have a number of live customer projects here, we're leveraging now the benefits of having both plasmid supply and vector manufacturing, albeit on different sites within a single source supplier.

Arsalan Arif (52:42):
All right, well congratulations on that and that sounds great. Andrew, thank you so much for sharing your time and expertise with us today. That's Andrew Frazer with Charles River Laboratories. It's all the time that we have for today. I want to thank everyone in the audience also for tuning in. Again, thank you to Charles River for sponsoring this discussion at Endpoints webinars. If you would like to re-watch this webinar or to share with your colleagues, a link will be available for on-demand viewing. I'm Arsalan Arif for Endpoints News. Thanks for joining us again, and we hope to see you at a future endpoints event. Thanks everybody.