Case Study: Advancements in Plasmid Manufacturing

Jeff Buguliskis (00:00):
Hello everyone and welcome to today's GEN webinar, which is entitled Advancements in Manufacturing for Rapid Delivery of Plasmid DNA Starting Materials. Our webinar today was made possible through sponsorship from Charles River Laboratories. I'll be your host for today's event. I'm Jeff Buguliskis, technical editor for GEN, which has been at the vanguard of the life science industry reporting on the latest news and the most innovative tools and technologies for over 40 years.

(00:45):
Over the past several years, we've seen some major improvements in development and manufacturer of advanced therapy products and there really are no signs that this continued success will begin to slow down anytime soon. With a steady increase in the number of products moving into late phase clinical studies and the potential to include more common disease indications with much higher patient numbers, there is an increased demand for high quality plasmid DNA supplies and greater pressure on worldwide manufacturing capacities.

(01:18):
Now the team at Charles River Labs believe they have an array of solutions that could be helpful to a host of research organizations. And our presenter for today's event is here to tell us more about them in just a few moments. But first, it's my pleasure to welcome Dr. Andrew Frazer, associate Director of Scientific Solutions and gene therapy at Charles River Laboratories, where he focuses on the plasmid DNA manufacturing platform and services offering. Andrew has a strong science industry background, where he has worked in diverse areas such as design, implementation and tech transfer of process use for the manufacturing of biocatalysts, small molecule APIs and biologics. And today Andrew is going to tell us about Charles River's plasmid DNA CDMO service offering, including platform screening and manufacturing approaches, improved supply chain management, in-house testing, and most recently, the launch of off the shelf plasmid products for viral vector manufacturing.

(02:24):
But before Andrew begins, I want to remind the audience that our Q&A session will begin right after the presentation ends. Now our Q&A session is much more fun and exciting if you get involved. So if you have a question for Andrew about his presentation or about some of the offerings at Charles River, all you need to do is type your question into the Ask A Question box and hit submit. And you could do this at any time during the presentation. You don't have to wait until the end and we'll try to get through as many of your questions as possible. All right, now I'm going to hand things over to Andrew so that we can get started and learn more about what's going on at Charles River.

Dr. Andrew Frazer (03:09):
So welcome everyone and thank you for taking the time to join me for this webinar where we'll look into advancements and manufacturing for the rapid delivery of plasmid DNA starting materials. So we'll begin with a brief introduction to Charles River before providing an overview of plasmid DNA and its role within advanced therapies, as well as some of the challenges and complexities that the industry is encountering within the advanced therapy space with regards to supply of plasmid DNA. We'll then talk about Charles River and how we are developing our plasmid CDMO service offering to address these challenges through the establishment of our XP DNA plasmid platform, the more recently the launch of our off the shelf plasmid products.

(03:59):
So Charles River was founded in 1947 and it has an extensive history and now a truly worldwide presence, with over 110 facilities in more than 20 countries. And while the company is most well-known for its capability in testing, Charles River now provides what could be described as a fully end-to-end service offering for drug development. And this goes all the way from research discovery, safety assessment and right through now to manufacture and testing. And we're very proud to have been directly involved with the development of more than 80% of novel FDA approved drugs in the last five years. And in recent times, about a third of these were within the cell and gene therapy field.

(04:54):
So hopefully that very brief introduction helps highlight the current position of Charles River within the wider advanced therapy space. And I'll now focus more on plasmid DNA and the role that it plays within advanced therapies. So plasmid is already in very high demand and the growing adoption and acceptance of advanced therapies utilizing plasmid DNA is driving further increased demand. I guess the relatively novel and complex nature of advanced therapy products is also creating challenges for suppliers and also consumers of plasmid products. And with that there are some clear opportunities for manufacturers to develop our approaches in response to help address some of these issues.

(05:44):
So technology around plasmid DNA and particularly its use within advanced therapies is firmly established and it plays a very major role within modern healthcare. The market for plasmid DNA has continued to grow very rapidly in recent years with market estimates in the range of 4 to $700 million in 2022. And the predicted growth of that market reflects the continued growth within the wider advanced therapy space. With compound annual growth rate, it's estimated over 20% and this leads to estimated market values in 2030 of over $2 billion.

(06:26):
The growing adoption and acceptance of advanced therapies utilizing plasmid DNA, both as a direct therapeutic product but also as a critical starting material for onward manufacturer of viral vectors, messenger RNA, transient proteins and other applications is driving increased demand. And as I mentioned previously, this in turn creates some challenges as specialist plasmid DNA manufacturers try to keep up with supply requirements.

(06:59):
So when we look at the application of plasmid DNA products within advanced therapies, we can see how the plasmid on its own can be used as a direct therapeutic product. So this would be the case for naked DNA therapies or plasmid vaccines. And this is shown in the top row in this slide. But importantly, it also plays a key role as a critical starting material for onward processing for many different advanced therapy applications. And with plasmid occupying this I guess, key foundational role within advanced therapies, it's critical to have reliable and cost-effective supply to deliver safe and effective treatments to patients.

(07:47):
So now on to some of the challenges within advanced therapies and some of the challenges that relate specifically to plasmid DNA. This slide shows some data taken from nature reviews and drug discovery looking into disruptions within the development of cell and gene therapies. And we can see in this example that unlike more established products like classical antibodies, we can see that lots of advanced therapies are having issues and complications that lead to delayed or discontinued regulatory submission and approval. And when we compare this directly with monoclonal antibodies in this example, we can see that cell and gene therapy products do experience a much higher rate of disruption for regulatory approval. And this ultimately drives up cost and time.

(08:38):
If we look at the root cause analysis, so section C on this slide, on the right-hand side, we can see that many of the issues stem from manufacturing comparability between clinical and then the commercial supply and it highlights a need to address CMC issues and also the requirement for more standardized approaches and stronger validation and continuity within manufacturing and testing when progressing from development through to clinical trials.

(09:12):
If we keep this in mind when we look at common sourcing and supply strategies for plasmid, it's clear that there are many areas where we can encounter these types of continuity issues. If we broadly group product developers or customers or consumers of plasmid DNA into three main types, we have academia, who are typically involved with product discovery and development and they often produce plasmids through their own research groups or sometimes dedicated specialist academic centers. And sometimes these academic centers also supply emerging biotech companies with early stage supply through development and preclinical stages. And we then have the first point of crossover with CDMOs, where essentially contract manufacturers supply academia, biotech and right through to large pharma. And it's generally when large pharma moves towards market supply for different license products that they consider or sometimes implement, bringing in in-house manufacturing and dedicated supply for plasmid DNA products.

(10:28):
If we then consider how this potentially looks as an example product, moving through discovery in clinical trials and then into market, we can see that there are many points of information data, physical material exchange between multiple organizations. This often includes academic groups, multiple CDMOs, due to the capacity restrictions that we talked about earlier. And then when we move to late stage, quite often there's a tech transfer element or even performing parallel development of commercial manufacturing processes, quite often for very well regulated and defined product at that stage. So changes are very, very difficult to make. So it is therefore very important that we do everything we can at an early stage with our plasmid supply to try and avoid issues that are common when transferring from one place to another and also think ahead as far as possible to make the transition from development and preclinical to clinical trials and then finally, market supply as easy as possible.

(11:39):
So to further look into sourcing considerations for plasmid DNA, developers have options on quality grade. This is based on a number of factors and it includes the clinical phase, the scale or volume requirements that are needed for a particular program and also compliance. And generally within the industry there are three quality grades that are used for different advanced therapy applications. So at the front end we have research grade plasma. This is mainly used for in vitro R&D or preclinical studies and it's generally produced in an R&D lab and comes with simplified documentation and also a reduced panel of testing to help reduce time and cost.

(12:28):
The next stage is high quality or HQ grade plasmid. This represents quite a big jump in quality from R&D grade and while it is sometimes used at very early stage, it's generally adopted for tox studies and is a critical starting material for onwards GMP vector production, generally within phase one and two clinic. And it can also be used for GMP messenger RNA production. In this case it does use fully traceable materials, much more comprehensive documentation and it's also produced in dedicated segregated production suites utilizing GMP principles.

(13:09):
Finally, we've got the highest quality grade and it's also the most costly and this is GMP plasmid. This can be used essentially for all stages of clinical supply, commercial viral vector manufacturing and DNA vaccines. And GMP products are obviously manufactured to the very highest possible standards in licensed GMP facilities, with fully comprehensive documentation, testing, QA oversight, and QP release.

(13:40):
So if you put yourself in the position of an advanced therapy developer, it does raise some questions over how you decide which creative plasmid to use and when. So it's fair to say that historically it has been challenging to clearly identify exact specifications and quality attributes for plasmids and the various applications. And this has made it difficult for both manufacturers of plasmids and also end users to make clear decisions in the development of drug products. So I'll not spend a huge amount of time going into detail on this slide, but the takeaway is that there is an evolving quality landscape for plasmid DNA and its use within advanced therapies.

(14:25):
That being said, the guidance has vastly improved and the 2021 EMA Q&A document and principles of GMP provides quite clear regulatory guidance on the quality standards for plasmid DNA used for all major advanced therapies. And essentially the end product, whether it be a naked plasmid, viral vector or modified T-cell, basically the closer it is to the patient, the greater the level of quality that's required. And it also highlights that GMP grade plasmid is not necessarily required when used as a starting material for advanced therapy products.

(15:03):
So to summarize then on challenges around plasmid DNA, with plasmid currently playing such a key role for so many different applications, it is in exceptionally high demand. And with that, there are some key challenges around it. These are centered around relatively straightforward things like production capacity and manufacturing capacity constraints, with slots at plasmid manufacturers often requiring booking many months in advance. An output of that is that many developers often need to navigate and manage complex and fragmented supply chain with a high risk of disruption when you move towards regulatory approval for advanced therapy products.

(15:52):
In addition, manufacturers and end users of plasmids need to clearly define on quality requirements for their specific application amidst this evolving regulatory landscape and then also leverage that into their supply chain to help address pressures on cost of goods and overall project complexity and costings for advanced therapy programs. So overall, there is a need for a phase and application appropriate plasmid solution that can provide reliable and also standardized production for a range of commonly used plasmids within the advanced therapy space.

(16:37):
So moving on then to address these challenges, Charles River launched our eXpDNA plasmid platform in January this year and this was set up with the focus of delivering firstly, exceptionally good timelines but also maintaining a very strong focus on process and product quality attributes. The eXpDNA platform is, it's a development of our longstanding plasmid service offering and it's based around three core capabilities. So the first is a plug and play toolbox approach where we can adopt various levels of screening and quality control checks to help mitigate the manufacturing risks associated with complex and challenging plasmid types that are commonly used within advanced therapy applications.

(17:31):
The second is a standardized platform manufacturing process and this allows streamlined supply chain and also documentation management. And lastly, we have the ability to implement a 100% in-house analytics for both cell bank and final plasmid product release testing. So by integrating these three capabilities, we can deliver what our industry leading batch timelines to our customers while maintaining that focus on quality. At the center of our eXpDNA plasmid platform are our plasmid facilities and last year Charles River established our dedicated Plasmid DNA Centers of Excellence in the UK. So our Keele site, which has been producing plasmids both for GMP and non GMP plasmid products for over 20 years now is now moved towards a dedicated GMP manufacturing site and it's currently undergoing expansion to improve capacity. Alderley Park is Charles River's newest CDMO manufacturing site and this opened in October last year. The site has three fully segregated processing streams for high quality plasmid with process facility and product quality specifications aligned to principles of GMP.

(18:57):
The development of these sites represents a very significant expansion to Charles River's CDMO manufacturing capability and it's also an important step towards addressing the aforementioned industry challenges and progressing the company's goal of improving the delivery of advanced therapy products.

(19:17):
So to go a little bit deeper now into the service offering, our Plug-n-Play Development Toolbox plays an important role within the eXpDNA platform. As anybody who's been involved in plasmid manufacturing knows that the cell bank is of critical importance in order to lay a solid foundation for any manufacturing program. And our cell banking process itself and also the testing around it have been in place for a long time. They're very well established. However, through many years delivering cell banks for plasmid products, we have developed an effective and also a robust approach to efficiently screen different plasmid types that are commonly used within advanced therapies. And this allows us to navigate many of the common issues that are associated with the generation of reliable cell banks.

(20:13):
Following an initial technical assessment of the plasmid sequence, our team will identify the type of construct and also any key features that are critical to its function. And this will then inform our approach for cell banking and dictate which, if any, additional screening or quality control measures that need to be included to ensure a very smooth progressions through manufacturing and into testing and release. Utilization of our ambr 250 system as a tool for early stage screening and pre-production evaluation can provide indication and process performance, productivity assessment and also yield prediction to ensure there are no surprises when we get to manufacture.

(20:59):
On top of that, we can implement pre banking, include selection screening steps to confirm plasmid integrity using methods like next generation sequencing, and these can all be used to effectively manage challenging plasmid types. So ITR containing plasmids, LTRs, long poly-A regions that are common to advanced therapy applications and this can all be done with relatively little impact on overall project cost and timelines.

(21:31):
The plasmid manufacturing production process itself also follows the same principles as our cell banking approach, so with a focus on rapid turnaround and also retention of event product quality, which has been brought together to help deliver reliable and right first time manufacturer of plasmid products. And the process itself is not specific to any one plasmid, but it has been designed in such a way that it can accommodate a wide range of different plasmid types without the requirement to perform lots of costly and time-consuming optimization work. One of the advancements within the eXpDNA platform is the development of both of our main production processes for high quality and GMP plasmids being able to utilize a fully single use process stream. And this includes dedicated single use pre-pack chromatography columns and materials and reagents that are completely free from animal origin.

(22:32):
So a fully single use process stream does obviously carry a number of benefits, particularly when operating our multi-product facilities, where we're often progressing parallel manufacturer for a range of different plasmid products for different customers. One of the main ones is that we avoid the time-consuming and costly equipment cleaning and residuals testing between batches. But also adopting a fully single use process stream effectively rolls out the possibility of product cross-contamination which can have very severe impacts and it can sometimes be very difficult to detect within some of these more complex cell and gene therapy programs.

(23:20):
While there are clearly advantages to running fully single use process, manufacturers have found recently that lead times for single use componentry can often often find itself in the critical path for manufacturing and project timelines. So while the manufacturers or suppliers of single use componentry are making their own improvements, it is something that Charles River recognized and as a response within the eXpDNA platform, we have put significant effort into standardizing our process and materials and also leveraging partnerships with our global suppliers. And it's a very important development in our platform for plasmid and it has allowed us to simplify our supply chain and also hold larger stocks of materials, which effectively allows us to initiate projects immediately on manufacturing and avoids any issues related to long lead time consumables.

(24:24):
So finally, one of the most important differentiators for our eXpDNA platform is the ability to utilize Charles Rivers testing powerhouse to perform a 100% of our characterization and release testing in-house. So our Plasmid DNA Centers of Excellence in Keele and Alderley Park are set up to feed directly into our cell and gene therapy manufacturing sites in the US. With all of these functions supported closely by our network of dedicated testing facilities and this effectively provides unprecedented access to testing capacity and capabilities. And it also allows us to closely coordinate our manufacturing activities and improve overall project timelines.

(25:11):
Now onto the most recent development within our plasmid service offering and one that's a compliment to our eXpDNA now platform and that's the launch of Charles River's off-the-shelf plasmid products. So when we first a little bit of background on plasmids in general within cell and gene therapy applications of viral vector manufacturer. If we look at the two main uses of plasmid DNA for viral vector manufacturing, we can see that the gene of interest, plasmids, are always therapy specific and these require custom manufacturer. However, the other plasmids that are used routinely for different viral vector products offer an opportunity to standardize. 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 utilized for Rep Cap plasmids, depending on the application. Equally, for lengthy viral vectors, three of the four plasmids, so the VSV-G, gag-pol and Rev are commonly used with different gene of interest plasmids.

(26:30):
So there is clearly an opportunity here to standardize on these commonly used plasmids and manufacture them in bulk amounts and make them available immediately off the shelf. Advantages to this type of approach are that it will help address capacity constraints. So one of the main challenges that we talked about earlier. That will help with lead times and also reduce the requirement for multiple repeat custom manufacturers of single plasmids. A result of that is that prices can 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 this helps avoid issues with under or over ordering and wastage.

(27:21):
If we look at this very simple example of a manufacturing campaign or to supply plasmids for onward AAV manufacturing, you can clearly see the benefit of Off-the-Shelf, helper and potentially, Rep Cap plasmid versus fully dedicated custom manufacturing. And these are mainly time and manufacturing capacity benefits. If you think about this a little bit further, you can also start to see a range of secondary benefits. So I mean, one of the main ones having been involved in lots of different plasmid products, is the reduction in complexity of the supply chain and also the ability to reorder plasmids at very short notice are significant advantages in today's manufacturing climate and they really shouldn't be underestimated.

(28:15):
In addition, the use of proven and standardized plasmids also has the potential to ease regulatory approval for the introduction of any follow on pipeline viral vector products in the longer term. So we are now very pleased to have launched our first off the shelf plasmid with our AAVP helper now available immediately in three quality grid. And this has all been set up to meet a range of customer requirements.

(28:47):
So it's available in R&D, HQ and GMP and you can check out our official press release on the Charles River website. In the future we obviously will be looking to expand on this part of the service offering and this slide summarizes our planned portfolio of Off-the-Shelf plasmid products. Also, the bottom shows some data from our AAV helper plasmid, which has been proven in the production of a range of different Rep Cap serotypes at various production scales for AAV manufacturer. So in addition to the AAV helper, we'll also shortly launch our lentiviral vector, Rev, gag-pole and VSV-G plasmids and these will all be available to our customers with full analytical packages and also in the same way as the helper, a range of phase appropriate quality grades for onward manufacturing.

(29:41):
Looking further down the line, we'll also aim to develop this service offering to include antibiotic versions of these plasmids based on our proprietary operator [inaudible 00:29:55] titration technology. And also in recognition of the interest in messenger RNA based therapies, we are exploring options for inclusion of generic back-bone plasmids for linearization.

(30:06):
So in summary, our eXpDNA plasmid platform coupled with the recent edition of the Off-the-Shelf plasmid products represents Charles River's initial steps forward in advancing manufacturing through the delivery of plasmid DNA and specifically plaid DNA starting materials to help expedite advanced therapy programs. We have been able to retain our scientific knowledge base and also our flexibility around different plasmid types by utilizing a toolbox approach to ensure reliable cell banking. We have adopted a standardized production process and leveraged our worldwide supply network for reliable processing and batch delivery and this is all underpinned by 100% in-house testing.

(30:58):
So this has allowed us to realize exceptional timelines for both HQ and GMP plasmids and also now the supply of Off-the-Shelf plasmid products. And as I mentioned previously is our aim to build on what we've started as we continue on the company's mission to support advanced therapy programs and also improve the delivery of these life-changing treatments to patients.

(31:26):
So thank you for taking the time to join the presentation and webinar. If you are interested in any of the Charles Rivers plasmid service offerings or also any of the wider elements that we mentioned in the presentation today, please don't hesitate to get in touch. Thank you.

Jeff Buguliskis (31:44):
Andrew, great presentation. Thanks so much for sharing with us some of the service offerings that Charles Rivers laboratory has and that could be really helpful to many members of our audience. So we appreciate that. Thanks again.

(31:57):
To the audience, Q&A is coming up in just a moment, so if you have questions for Andrew about his presentation, all you need to do is type them into the ask a question box and hit submit. Okay, without any further ado, let's get onto the Q&A. Bear with us for just a moment as we make the transition and we're going to try to get through as many of your questions as we can.

(32:22):
All right everyone, thanks so much for joining us for the Q&A session. Andrew, again, thanks for great presentation, really appreciate you sharing all that info with us. We have a couple questions here from the audience, so let's get to them, try to get through as many of them as we can. The first question we have, an audience member would like to know or says, "You mentioned that Charles River has plasmid viral vector manufacturing and testing capabilities. Can you comment on the benefits of being a single source provider versus working with multiple CMOs?"

Dr. Andrew Frazer (32:54):
Yeah, absolutely. So I think we kind of touched on it in the presentation and I think the first thing probably to say is that every project and program is different, but like I say, as we touched on in the presentation for advanced therapies in particular, we kind of saw some of the data around increased issues and complications with CMC and disruption for approval. So that accompanied with often very complex supply chains, there's clearly significant scope to improve. So the supply chain kind of example or I guess, generic example for plasmid that I've presented, you could extrapolate that out to cover viral vector manufacturing and beyond for cell therapy based products.

(33:47):
So quite often developers find themselves working with three or four or even more service providers and manufacturers and beside simple efficiencies and communication, just simple scheduling of manufacturing activities and testing and the communication around that, removing the need to physically move material between different providers on its own can easily take months off a manufacturing timeline.

(34:16):
And in addition to that, I think having a single multidisciplinary team all pulling in the same direction under single management definitely supports a much more effective and timely decision making and problem solving process if that's required. And equally, it allows project managers full freedom to align manufacturing timelines and influence scheduling and optimize that to coordinate with ongoing kind of study activities.

(34:52):
So I think to summarize, it's probably difficult to put numbers on it because like I say, everything particularly in advanced therapies, it's quite bespoke. Everything's quite new. We are in the early stages but clearly there are huge advantages and at Charles River that's something that we're really focused on trying to deliver by having that end to end kind of service provision.

Jeff Buguliskis (35:21):
Alrighty, thank you for that, Andrew. Next question we have comes from John who says, "Besides lead time, what are some other advantages of the Off-the-Shelf plasmids?"

Dr. Andrew Frazer (35:35):
Yeah, so Off-the-Shelf plasmids are not kind of new to the industry, it's something that is new to Charles River and what we're doing within our platform, but lead time is obviously a key benefit. But I think on top of that, if we have these commonly used plasmids available, it does help to reduce pressure as a manufacturer. So on our manufacturing capacity and our testing capacity. So that is a real advantage versus fully custom supply where you need multiple plasmids for example, for onward viral vector manufacturer. On top of that, I think being able to generate Off-the-Shelf plasmids in larger bulk quantities also allows the prices to come down and like we said in the presentation, customers can kind of plan ahead and place orders on a gram versus a batch basis and avoid issues with wastage.

(36:32):
I guess on top of that, if we kind of look for further advantages, things happen in manufacturing, particularly through early development, you can easily encounter failures, you can have technical issues with process or testing. Quite often, reworks need to happen. So actually having that option and that ability to reorder and restock your plasmid supply at short notice is quite a big benefit and an advantage versus having to go out and secure manufacturing capacity and run dedicated manufacturing, which can sometimes take a number of months and that is a huge benefit.

(37:19):
Again, on top of that, I think in the longer term, if we do see widespread adoption of standardized plasmids, it could help with regulatory approval of new products and particularly if a developer has a product that they've already progressed through regulatory approval, it'll definitely help support bringing on any follow on pipeline gene therapy products.

Jeff Buguliskis (37:46):
All right, great. That makes sense. Thank you for that. Next question we have is from Vanessa who says, "Are there differences in regulatory guidance for plasma DNA between the US and EU and are your UK manufacturing sites compliant with FDA regulations?"

Dr. Andrew Frazer (38:05):
Yeah, it's great question. It's one that comes up quite a bit. So I think firstly, I would say that while our UK GMP manufacturing is performed under license from the MHRA, we do definitely have compliance with FDA regulations and we have supplied plasmid products for all stages of clinic, right through to commercial. I think with regards to differences in regulatory guidance, if we look at what the FDA have done in a similar way to what the MA have done around principles of GMP and clarifying that line between GMP grade and intermediate grade plasmid, they have come out and said, similar to what the MA said, that you don't require that plasmid starting material for onward vector manufacturer is generated under strict GMP. It doesn't need to be treated in the same way as a plasmid that used for drug product or drug substance for example, or in the case of a direct injectable.

(39:13):
And they also did state that, similar again to the MA article around principles of GMP, that while plasmids do not necessarily need to be GMP, many of the attributes of GMP product need to be implemented. So I would say in that regard it's all about taking a risk-based approach with these things and I would say that the guidance between the US and the EU are pretty well aligned.

Jeff Buguliskis (39:41):
Alrighty, thank you for that, Andrew. Next question we have comes from Ramesh who says, "There seems to be a trend towards increased quality standards for plasmid DNA. Do you expect that testing specifications will follow a similar trend? For example, will specifications that are acceptable today become unacceptable later?"

Dr. Andrew Frazer (40:05):
Yeah, so I can definitely see the level of qualification used for various methods of testing increasing and becoming more aligned to what we see for GMP products. And again, this kind of goes hand in hand with what we talked about around risk-based approach for justifications of principles of GMP rather than full GMP for particular applications. I think specifications on testing are a little bit more tricky. There's a very useful Biopharm article that focuses on release specs for plasmid DNA that was brought recently. And it is clear that there are currently lots of variability around what manufacturers offer and even well-established specifications like plasmid super coiling are quite difficult to standardize and dictate.

(41:00):
So for example, the consensus in the industry is that higher percentage super coiling delivers better transect efficiency and productivity. But I would say in reality it's probably not so clear cut and we probably need to develop more of a focus on plasmid and application dependent data so that in the longer term, as advanced therapies become more well-defined and standardized then so too will the specifications around starting materials, like plasmid.

Jeff Buguliskis (41:37):
Alrighty, thank you for that. Next question we have is from Joe who says, "Manufacturing production scale seem to be relatively small compared to monoclonal antibody production for example. Why do you think that is?"

Dr. Andrew Frazer (41:54):
Yeah, I think it's true, particularly for CDMOs, or some CDMOs and I think some of it is related to what I talked about in the presentation around widespread adoption of single use technology and manufacturing. And also the requirement for CDMOs to operate multi-product manufacturing within their facilities. And we talked about some of the advantages of this approach, but I guess the disadvantages with single use technology is that when it comes to scale, there is a technology limitation associated with it. The largest kind of single use microbial bioreactors at the moment are kind of 3 to 500 liters. So at that scale it does work well and it definitely provides flexibility for onward indications with relatively small patient numbers or plasmid requirements within the kind of a 100 gram quantities.

(43:06):
But for products with much larger patient populations like hemophilia or DNA vaccines where you're looking at direct injectable products, then you're looking at tens of kilos of plasmid per year, or even more. This is typically where you'd see a swap over to dedicated facilities in-house manufacturing and more of the traditional stainless steel facilities that can operate at much larger scales.

Jeff Buguliskis (43:37):
Alrighty, thank you for that, Andrew. It looks like we have time for one last question. Gregor asks, "What is your largest capacity GMP plasmid DNA production quantity for customer plasmids?"

Dr. Andrew Frazer (43:52):
So yeah, so currently at our Keele facility in the UK we have 50 liter upstream production for plasmid DNA, and we can, use that for multiple different plasmid types. Obviously actual plasmid produced is highly dependent on plasmid type and there is a range associated with that. So yeah, 50 liter is our current largest capacity that we offer for GMP. We are planning to implement 300 liter processing in the future. So that's something that is definitely in the pipeline for us and that will be to GMP standard as well so we can accommodate those larger requirements.

Jeff Buguliskis (44:37):
All right, great. Thank you, Andrew. And with that we've come to the end of our webinar. So I'll like to remind everyone that the webinar will be archived on the GEN website that's at genengnews.com, for up to a year. So if you missed any parts of it, you can watch it again, or feel free to forward the link to any of your friends and colleagues who you think might be interested in the content.

(45:00):
I'd like to thank Andrew again for his very informative presentation for great Q&A and you the audience for your attention and very thoughtful questions and a very special thanks to Charles Rivers Labs for sponsoring this webinar. Hopefully we'll see again at another GEN webinar in the near future. Goodbye for now.