Oxford Biomedica plc (OXB) Earnings Call Transcript & Summary

Good day to everyone joining us, and welcome to today's xtalks webinar. Today's talk is entitled Fourth generation Lentiviral vectors and improved gene delivery system. My name is Sonia Hunte, and it's my pleasure to be your xtalks moderator for today. Today's webinar will run for approximately 60 minutes. This presentation includes a Q&A session with our speakers. This webinar is designed to be interactive, and webidars work best when you're involved. So please feel free to submit questions and comments for our speakers throughout the presentation by using the questions Chatbox and we'll try to attend to your questions during the Q&A session. [Operator Instructions] At this time, all participants are in listen-only mode. Please note that this event will be recorded and made available for streaming on xtalks.com. At this point in time, I'd like to thank Oxford Biomedica who developed the content for this presentation. Oxford Biomedica is a quality and innovation-led viral vector CDMO with a mission to enable its clients to deliver life-changing therapies to patients around the world. One of the original pioneers in cell and gene therapy, Oxford Biomedica has more than 25 years of experience in viral vectors, the driving force behind the majority of gene therapies. Oxford Biomedica collaborates with some of the world's most innovative pharmaceutical and biotechnology companies, providing bio vector development and manufacturing. Oxford Biomedica's world-class capabilities span from early-stage development to commercialization. These capabilities are supported by robust quality assurance systems, analytical methods and depth of regulatory expertise. Oxford Biomedica is headquartered in Oxford U.K. It has locations across Oxfordshire U.K. and near Boston, Massachusetts, United States of America. Now it's my pleasure to introduce you to your speakers for today's webinar. And first, I'd like to introduce you to Nick Clarkson. He is the Vice President, Head of Platform Research at Oxford Biomedica which drives innovation of Austria Medical's core lending factor platform through vector design and improvement, generation of packaging and producer cell lines, small-scale process development, computer-aided biology and development of core analytics. The group's mission is to keep the Lentivector platform at the cutting edge of gene therapy and to ensure applicability rather, to the needs of a wider cell and gene therapy community. Nick obtained his PhD in molecular immunology from the University of Oxford. And next, it's my pleasure to introduce you to Dan Farley. He is the Senior Director of the Vector Engineering Group. Well, there's Nick. Hi Nick. Now it's my pleasure to introduce to Dan, and where is Dan? Dan Farley is the Senior Director of the Vector Engineering Group at Oxford BioMedica. He joined the company in 2004, having completed a PhD at Warwick University working on adenoviral vectors and Soundline development. The Vector Engineering Group was founded in 2013 to improve OXB's lentiviral vector platform and technology for cell line development, which later expanded to vector agnostic technologies encompassing AAVs and adenoviral vectors. Now it's my pleasure to pass the mic and the controls over to our first speaker, and that is Nick. So Nick, when you're ready. You may begin.

Thank you, Sonia. It's a pleasure to be here, and thank you all for joining us for this xtalks' webinar where we're going to talk a little bit about TetraVecta and Lentiviral vectors in general. What we really hope that you're going to learn today is some of the advantages of using Lentiviral vectors in cell and gene therapy. How you can improve titers, and I'm going to show you some of our normal technologies that we've developed. We're also going to talk a little bit about some of the important aspects that can derail or delay product development. And this can be anywhere from particle quality interactions of your gene of interest that may affect processing and use. Also, some of the big questions like transgene cargo capacity because it's always an issue in cell and gene therapy and also talk a bit about the potency and this is potency of gene of interest expression in your host cell and how it interacts with the host. And finally, we'll hopefully learn some of the novel tools, including our fourth-generation Lentiviral vectors that are actually being designed to address some of these needs and issues that are encountered. So this is the agenda today. I'm going to give a brief overview of Oxford BioMedica. And then introduction to Lentiviral vectors, then I'll hand over to Dan, who's going to take you through the third generation and fourth-generation LVs, and we're going to finish with a summary kind of question-and-answer session. But just to begin with, we have a little poll question. So over to you, Sonia.

I'm going to jump in right now and launch that poll question. And just let the audience know that this poll question is done in real time. So your participation is strongly encouraged and very much appreciated. Now I'll give you about 45 seconds to complete this simple poll question. So please go ahead and pick select any of the below. The question we had is do associate Oxford Biomedica OXB with specific capabilities or expertise in Lentiviral vectors. So please go ahead and select from the below. Is it yes, OXB solely specializes in LV or yes, OXB specializes in LV and other vector types or is it now? So please go ahead and make your selection. Okay. That looks like the majority of our audience have cast their votes, so I do appreciate that. I see that there's some other members there that have not. So I'll leave it open for a few more seconds and hoping that you will be encouraged to participate with us in this poll question. Perfect. That looks like it's the majority of everyone now. Thank you so much, everyone, for participating in the poll question. As I said before, all you have to do is click on any of the options below Okay? And that looks like is everyone. Thank you so much. I'm now going to close the polls, and let's take a look at the results. Okay. So our audience members have made their selections. And here are the results. We have 25% of our audience that, yes. They said that OXB solely specialized in LV for what they thought. And then 68% shows this one here. Yes, OXB specializes in LV and other vector types while 7% said no. So thank you, everyone, for participating in poll question #1. Now back to you, Nick.

Thank you, Sonia. I mean that's really interesting. We've originally started off to specializing the LV, and recently, we moved into other vector types. So it sounds really interesting to see that the message is actually getting out there -- so tell me, if you can see my screen? Is it moving okay?

Yes, it is.

Correct. So I'm just going to give a brief introduction to talk to Biomedica. Sonia have some very nice words about us. But we've been around for 25 years, and we have a broad range of experience in viral vectors. Everywhere from lentiviral vectors, which we initially started with to have no associated viruses and also adeno manufacturing. And to date, we've made over 340 successful GMP batches in the last 10 years. This is mainly due to our client base. We are -- internationally, we have 24 global clients, 41 partner programs and our Lentiviral vector platform is approved with products in over 40 countries. What really makes us successful in it. It's our end-to-end services for all the vector types, everywhere from construct and plasma design, analytical development, all the way through process development, manufacturing, QA release and stability studies. What's really underpinning this is our proprietary platform that we can offer an AAV lentiviral vector and adeno, this is really backed up with our quality systems expertise in our state-of-the-art facilities. And in addition to our proprietary platform, we also have the ability to take transfer in other platforms from outside of the organization. So really, our success is based on our innovation, and we have what we call our innovation engine, and this is the multiple breakthroughs, particularly in the LV technologies and now in the AAV field. We spent a lot of time looking at selling vector engineering. This is really gave birth to our next-generation antiviral vectors that you're going to hear about shortly, the fourth gen factors. We also produce cell lines, which is something that we've really excelled in the recent years. We innovate a lot in a large scale process production, and this is net new platform using perfusion for an lentiviral vector. And this is defusing with certain enhances that we're going to talk about. This boosts your titer and quality. So we're also very keen on the analytics and data. So we maximize our use of data and integrate it, and this leads to us using a lot of design of experiments to optimize plasmid ratios and transduction and other prompters. And we use things like proteomics and transcriptomics to characterize our viral vectors and being very complex biological entities, you need some quite advanced techniques to understand them and characterize them. And then moving forward, we actually have automated a lot of our assays going into Q3 release, which gives you the precision and consistency. This required for active reduction for human yes. So we do have a second poll question. This really does allow us to understand the market and what how people see it. So over to you, Sonia for more question, please.

Certainly. Okay. Here is this everyone poll question #2. And there you go. It's all about you, which industry segment do you represent? We have a couple of options below that you can choose from. So please go ahead and make your selection. Is it work in biotech or pharmaceutical company. I am Academia. I work for a CDMO, CRO or service provider, I am an investor or none of the above applies to me. So please go ahead and make your selection everyone's quick this time, we got the majority of our audience have cast their votes already. So thank you very much for that. I'll leave it open for a few more seconds as we have a few more people there who are just making their selection. Okay. That looks like it's the majority of everyone. Thank you, everyone, for participating in pull question number 2. Let's take a look and see who's in our audience. Okay. So in our audience, 23% that I work in a biotech or pharmaceutical company, while 7% that I am from academia, with 51%, say, I work from a CDMO or CRO or a service provider and then 11% or I am an investor and 8% are net of the above applies to me. Those -- so thank you, everyone, for participating in poll question #2. Now back to Nick.

Thank you, Sonia. Again, that's very, very quite interesting. Hopefully, if you join this webinar that sound you a bit of [indiscernible] whether you're from academia, from CEO industry or even investors that we focus on. You'll find it interesting. So just to check the slides are changing?

Yes.

So I'm going to give you a brief introduction to the Lentiviral vectors. Now Lentiviral vectors, they're well known in the industry, and they are particularly good at delivering large payloads and they have an excellent safety profile and long-term gene expression. Now the most commonly used Lentiviral vectors for gene therapy is typically based on an HIV 1, which is what we most commonly use in our platform, and it has the ability to stay and integrate into the host genome. It's particularly good because it's nonpatent, which Dan will talk about in a minute. And there is a lack of pre-existing immunity in the general population. It can deliver genes to both dividing and nondividing cells. It actually does it in a very efficient way, which it enters the cell, it protects the cargo to protect the RNA limited to the nucleus, you only need one integration event that will happen, and it's fairly integrated into genome and then you get the effect of the gene expression. And the fact that it does this really means that it has always a lot of the innate responses. It avoids a lot of new creators and base modifications that can happen as part of post natural defense against viral vector. So it makes some really useful permanent modification of sales. The other advantage of delivering up to 10 kb of transgene capacity. And it now actually being considered for in vivo high-dose applications, and this has been made possible through innovation in [indiscernible] targeting traditionally as Lentiviral vectors have vector of renewed [indiscernible]. So saying they might be adopted in the cell and gene therapy and there's 9 commercial products to date. And they actually make up about 43% of ongoing clinical trials in cell and gene therapy. And I mean, the biggest market really is in oncology. Everyone probably aware of CAR-T and the amazing results that have been achieved by using modified T cells in oncology. And the majority of products now are moving forward in the CAR-T based on Lentiviral vectors gene delivery. So I'm now going to hand over to Dan, and Dan will take you through some of the aspects of the third-generation Lentiviral vectors and then talk to you about some of the improvements made and the fourth generation vectors. So over to you Dan.

Okay. Thanks, Nick. So as Nick said, I'm going to just give you a little bit of introduction in terms of state of the art and then we'll move on to TetraVecta and what that means. Okay. So this slide just gives you an overview of the generation lentiviral vector components. So it's a full plasmid system. And I'm highlighting here the kind of main takeaways from this, right? So focusing on the genome plasmid, third generation vectors are driven by a powerful promoter that drives the vector genomic RNA they get packaged. And this is important from previous generations because that happens in a way that's independent of the auxiliary protein called Tat, which we've seen as an important safety feature. Just highlighting a couple of key elements in there. So sequences that are retained in parts of the sequence the kind of legacy sequences from HIV from which the vector is derived and that includes the packaging region, which contains this major splice donor site and the reasons why we're highlighting that will become obvious data. The red response element of this element is necessary to be acted upon by REV, which allows you to generate patchable RNA. And third generation vectors has also employed what called self-inactivating LTR and this removed promoter activity and was also seen as an important safety step. The actual transgene cassette relies on the polythene latency sequence that terminate transcription of the gene interest mRNA and the target cells, and as we'll go on to later on, we'll talk about the relative strengths of these sequences. And in production, it typically include a backup [ polyon ] reasons for that will become obvious later. The BPRE, which is an element, it typically included to increase especially in target cells is also one of the main features of third gen when we employee a mutated version of that. Okay? So -- and then moving on to packaging plasmids. We have three of those components. So that includes the Gag-Pol cassette which we cure and optimize, and that has the effect of both minimizing homology between some of these components to greatly reduce the ability of these components to recombine and potentially generate something that's infectious. The other consequence of that is the The other consequence of that is the Gag-Pol plasmid is no longer REV dependent. So typically, most Gag-Pol -- historically, most Gag-Pol also have an RE element as they do in the genome, and that's dependent on REV. So [indiscernible] has genome system. And again, the reason for highlighting those will become of this data. And then we're obviously go expressing the growth cassette and then you have the ability to see the site, lending of our vector particles by supplying any kind of viral spike pricing, but typically, the feel that has used [indiscernible]. Okay. So we wanted to just pause briefly before we moved on to TetraVecta to talk about some of the technology that we have developed to just increase the output of lentiviral vectors in general. One of those is called U1 technology. And this is essentially U1 sRNA molecule, which is a noncoding RNA, which is normally involved in splicing in all of our cells. And what we've done here is we've taken part of the sequence here that is on that U1 sRNA. And instead of being able to involve an splicing, it can now bind to the beta RNA. And in doing that, in coproduction with the other components we just described, we see kind of a dose response. So the more of that plasma we put in with the type get and we see anywhere between the sort of 2 and sometimes we see up to tenfold increase in titer. And the main reason for that is because we are stabilizing the pool of packageable RNA in the cells, and therefore, you're actually able to fill more of the particles that you're generating. And that has the effect of improving P to E ratio. Here that the lower than number of particles mean that more of those particles are active. And so we can talk about this technology is being used by some of our clients in GMP to a great benefit as of today. The second technology we want to talk about is our U2 technology, which is a class of protein kinase. And we've essentially found that these are able to give the cells a bit of a kick during production and increase it's expression from the promoters that we're employing to drive expression of lot of those components. And this particular version, which is called Engine of the Alginate, we can see that we're able to see some -- again, some quite significant increases in titer and again, also an improvement in this P to E ratio. So the specific activity the parcels we're generating is improving. And we have 2 technologies together to get combined effects and quite a lot to show that these in users are cleared and in case the U2 down on the [indiscernible] level. And so these 2 technologies, we recommend also combining with our TetraVecta system as well. Okay. So I'm going to change that and move on to TetraVecta System. But before we do that, we thought that we would do our third poll. So I'm going to hand over back over to Sonia to take us through that.

Okay. Dan i will run our last poll question now, and here it is, everyone. What do you consider to be the key desired attribute for future lentiviral vector systems? So please go ahead and select just one of the below I'll give you about 5 seconds to complete this poll question. And just like before we'll close the polls, and we'll take a look at the results. Is it the highest titer? Highest quality and safety? Larger packaging capacity enhanced potency and effectivity? Gene expression or easier manufacturing? So please go ahead and make your selection. Okay. Dan, it looks like we've got the majority of our audience have casted their votes. But as you know, I have to leave it open for a few more seconds. For those of you who have not selected if you did make the selection now. I'll leave it up the first few more seconds, and then we'll close the polls and get back to the presentation with Dan. Okay. That looks like it's everyone. So I'm now going to close the polls, and let's take a look at the results. Okay. So here are the results, Dan, we have in our audience, 13% is the highest titer, than we have 51%, 51% of our audience had highest quality and safety while 8% said larger packaging capacity and then 15% for enhanced potency and then 13% said easier manufacturing. So thank you, everyone, for participating in our last poll question. Now back to you, Dan.

Okay. That's interesting. We had an interesting internal discussion about whether, which of those we thought would be top. So it's interesting to see quite an interesting mix. So -- and I think that kind of leads us nicely into this line. So really, we wanted to kind of highlight some of the experiences we've had in developing lentiviral vector manufacturing processes for our clients and the also for ourselves. And sometimes, this consumer has been like a black box. So we have a gene of interest. Select made some variances [indiscernible] optimization. And then we may certainly be thinking great let's just get some LentiVector made is all going to work out fine and we'll be often away in a clinical trial. And really, we want to highlight that obviously an aspect of that as long as we get the right yields, everything is fine. And it's been our experience that the transgene and LentiVector design can have a significant impact on the success steps through this pathway to actually realizing a batch that you're happy with and going into whatever kind of trial we want to perform. And so taking some of these one by one, right from the word go, I guess one of the questions we'll be asking people have you already been able to onboard your favorite candidate gene of interest cassette? Well, you already have to make a compromise because you haven't had an issue at the moment you onboarded onto the genome, you didn't make enough titer. One general thing we would ask people is, do you know the source of your LentiVector cassette. We have some experience with some very old legacy G&As with clients coming in and not quite understanding what's happening to that cassette. We're also aware of widely available lenti geno plasmids that have significant open reading frames from HIV left in some of these backbone sequences. So that's a good question to ask yourself. Do you understand the vector you have in your hands? Then there are other questions. Sequence optimization to be cured and optimize? What about splice size? Repetitive sequences, that maybe the size of payload. I was having to compromise on what we on board because the prices tend to go down the payload. And then also probably one of the more significant aspect, is gene of interest expressed in production? And is that impacting on your upstream titer? Is that gene of interest a secreted or membrane protein. So is it ending up in the state that's going into your downstream process? And then the potentially knock on effect what are some of these consequences with quality of the final product. Have you actually got a protein in the final product will come back and touch on that. And obviously, there's all of this above can potentially impact on the success of this pathway. And so we're arguing for a kind of LV design-centric to you, spending a bit of time here, getting things right before you move through this pathway. So now we're going to go on to talk about how we think, et cetera, that simplifies what potentially is completed process. Okay. So what we've really done with TetraVecta is sort of playing on ideas being a fourth gen and four aspects. So most people agree, it sounds like from that poll that most people are, believe that all these aspects are important. And so we're going to take you through these 4 technologies that will address one or all of these. So firstly, that's the TKO genome, which has performed really part of the core of what TetraVecta is. We're going to talk about the trip system and how that works better in TetraVecta System. And we're going to talk about MaxPass, which is more perhaps sort of capacity aspect. And then we're going to talk, finally talk about our [indiscernible] which is really not kind of safety and patent expression in the target. Okay. So the first really technical slide on TKO, as I said, this is really the core of TetraVecta. And so taking you through here we've got a schematic showing of what you kind of RNA is actually generated for the third-generation expression cassette during production. And so obviously, ideally, the mRNA which you want to package and usually if your internal promoter is active, you will also get some expression translation of your gene of interest. But what we -- and also, you can find in the literature as [indiscernible] that there is a lots of [indiscernible] splicing happening. And that is happening from this milestone that's embedded in the packaging signal of that that's derived from HIV. And so what happens is you get bypassing of the [indiscernible] RE element even in the presence of REV. So the [indiscernible] favorable finds the element here and stop this from happening. And so the consequences are that you make this truncated RNA and you will get in gene expression from here. And so this also goes some way to explain why perhaps some people might have a tissue-specific promoter that they don't think adaptive in the base cell lines, say, they might take that cassette from an adeno or an AAV because et put in at and suddenly they're tissue-specific promoter lights up the cell. And really, the reason for that is splicing activity. And so we can see that in this RT-PCR. We can see that we have total RNA here in the cell and what you can actually also detect in the [indiscernible]. So we can actually detect the supplies for us in [indiscernible]. Okay. And so what we've essentially done is we've been able to mutate and to ablate all this splicing activity from our TKO genome. And that leads to very clean production of full-length packageable RNA in your production cells and that obviously translates to a single species, is being present within the vector particles. And so during the course of our study, we found in doing this, you actually reduced the titers. And so we're able to rescue those solutions by rescuing the titers with dependent version of our [indiscernible] are you on that I've described previously. Okay. So I'm going to move on to talking about Trip System. So our TRiP System was actually developed a number of years ago now. And it's a translational repression system that starts to expression of transgene only in production. And that happens by co-expression of a protein called TRAP. And that bonds to its target sequence in the RNA that's placed immediate upstream the primary starter protein. And so you stop the ability of that mRNA to be translated. And so what we're able to do here is to stop the side effects of your keen of interest, and that might be some obvious side effects like cell viability or perhaps some less predictive things like on assembly, which might impact on, say, your upstream, your output, the amount of vector is actually coming out of yourself. Even if that doesn't impact on titers, as I said before, you can have issues with the origin of interest being shed into the feed or even actually embedded within your vector particle and that might have some consequences downstream in downstream purification. Okay? So we've generated sequences now that can kind of plug and play a lot of different settings, and we've demonstrated that for both Lenti across other Larva vectors as well. Okay. So the TRiP System was developed really to kind of park a lot of these issues with sort of specific to the gene of interest you're implying. Okay. And then so what the issue is with the third gen vectors is that some of those splice vasogenic RNA that can encode transgene are poorly repressed in the Trip System. So what we're able to do is partner both the Trip System into our TKO genome, and we're out to get a lot better levels of repression. And that's leading to significant increases in titer to where we know that the transgene is having a problem. So in this case, the primary antigen receptor, gene of interest for CAR-T, where we know the titles are very low. And then by employing TRiP on a [indiscernible] gene, we're able to see both on the expression of the actual CAR, which has demonstrated the Trip doesn't have an impact on your target case transport there. And integration in another report on the amount of debt are producing it. Just one little comment here is that if to say titers are a relative term. Maybe we can touch on this in the Q&A. So because people use different assays to turn in their sites, it's not very helpful to try and compare between different platforms that we probably should pick up in Q&A. And then, so this is what's called the Vector Block. This is showing the level of protein, both in the production cells of bulk here. So in this case, when we're employing on a promoter, this is poorly refreshed, and in our [indiscernible ] genome, we're able to see really nice lever of repression relative to the [indiscernible] . And we're also able to demonstrate that this stops CAR from entering into your particles. And so perhaps for something like car, this could be really important. So for example, if your CAR perhaps might target your lentivector to a B cell, you might not want that to happen because over expression of your CAR in a B cell, are your target [indiscernible] CAR-T therapy might downregulate the antigen. And so you might get some kind of pseudo negative relapse in that indication. So TRiP is really nice way of really kind of down regulating team of interest protein is in your breath. Okay a switch tack again and talk about MAXPASS. So I realize this is a little bit complicated this side. But what we really wanted to do is to show you the kind of 2 flavors of genomes that we are offering under the TetraVecta platform banner. So I previously talked about what we're calling TKOs. So because of this mutation, we require the U1 enhancer during production to give us maximum titers. And during the course of our studies, we were looking ways of being able to make TKO genomes without U1. And essentially, the way we've done that is to replace the RA, which is normally acted on by REV with a synthetic vector in from what we call a vectoring from them. So what's happening here is that we're just splicing out a little synthetic in from and that's leading to stabilization of the vetting the RNA okay? So the reason, so there are [indiscernible] ways in which we're applying this. So the caveat with MAXPASS genomes is because we're stimulating splicing, it means that your gene of interest or your case, you're trying to deliver to patients can't have an intron because those will be lost. And so the reason we are offering these 2 options is if you really want an intron in your cassette, you would use this TKO genome. And if you're not bothered you would probably go for a MAXPASS. And to be honest, for indications where you're trying to cram as much possible into your cassette, you're probably already thinking about minimizing some of the [indiscernible] features like in from to try and squeeze in as much into payload, okay? So the other thing I should say is because we're using, going back to what we're saying earlier, we use a code we use a GAG [indiscernible] as polar independent. So that means we don't actually need REV on our system. And so for MAXPASS, we're able to minimize down to 3 plasmids and we think that that will significantly help the optimization around production of MAXPASS [indiscernible]. So here's a little bit of initial data, which is interesting, showing that when we can generate a bit more space in the backbone, we're able to see some simulation in titers. And so obviously, the [indiscernible] itself doesn't contribute to splicing, okay. So we've been able to remove the RE. We've also been able to remove some other aspects of the patches to give us about [indiscernible] won venture space. Okay. Okay. And then I'm going to talk, take you through a couple of slides on our [indiscernible]. [indiscernible] stands for Sequence Upgraded Poly LTRs. So what, going back to our sort of comparison to third gen. So the self-inactivating feature that was percent of those genes. It resulted in deleting the promoter activity sequence, but it also had the effect of deleting poly sequences enhancers that we do want, and so the consequence of that is that the poly [indiscernible] that are used by the transgene cassette are weak. And so there is reading out this transcription of reading out of an integrated cassette and there's also potentially transcriptional read in cassette. There may be some position effects that happen on your [indiscernible] that you might not want. Okay. And obviously, if your [indiscernible] has a major splice donor, then it might part kind of slicing out either into the cassette or outside. And there's been some applications on it at in the literature. Okay. So the [indiscernible] we've essentially put back in enhanced elements in a way in which we've not put back in the promoter disease, so still a SIM in a traditional sense. But it harvest these much, much improved [indiscernible] sequences. And we've also been able to provide transmission installation on the bottom strand we're able to stop any transcription coming through on the bottoms trend as well. We've seen a stimulation in the level of expression as well, which I'll go on to hear you now. So you're going to get from left to right. So here's just a compact bit of data showing a comparison where we've just got a like a list phrase report here in a production cell showing what happens if you just don't have any termination which is control. That compares to SIM LTR,and then that's now comparing to our optimize. So we're seeing a pretty significant increase in [indiscernible] and [indiscernible]. And then when we look at an integrated NT comparing both third and [indiscernible] combo, we're again seeing something like a 50-fold reduction in transcriptional read-in from cellular promoted into our lenti cassette Okay? And then on the third panel here, we're showing some data across different cell lines using different promoters showing that we're starting to see a consistent, albeit modest, but probably helpful improvement in transgene expression somewhere in the region of 2 to 3 [indiscernible] average. Okay. And then this is kind of putting it all together, just a summary slide comparing, putting all these things together, as I said, these are kind of is the 4 aspects that we can kind of potentially put in a modular gene [indiscernible]. You want depending on maybe whether your gene of interest is not giving you an issue or not? It really just kind of give a summary of what I've just been talking about. So we populate this advertising. We no longer see these large products in particles. And we have the ability to be [indiscernible] also stock transgene interest being embedded within the feed. So we think that this would potentially translate to cost savings through downstream process and and potentially shorten that development time as well. And as I said, just to give you an idea of what the build your can do in terms of [indiscernible] installation? Okay. And then coming on to really what my last technical slide. So particularly for chimeric antigen receptor [indiscernible] cellular lines, it's been found that really in transient that this is typical for those card not to show any kind of acute toxicity problems. You're only manufacturing over a number of days. That's not to say that we haven't found particular CARs where the trip system benefits. But I would say, generally, it's not impossible to make CARs in transient. But when you move to a stable cell-line setting where you're components are integrated into your producer cell line, you're then talking about having to keep those sales going for a long, long time through the banking process and ultimately what they're actually building up a seed train to into GMP manufacturing. What we really find is that these CARs are actually quite chronically toxic. And that's had real implications on the ability to generate produces our lines for CARs. And so we've been able to combine current aspects, the main [indiscernible] and the TRiP System to really shut down or minimize expression of CAR. And that means really you get a bigger pool of sales coming through to screening and you're much more likely to pull out claims that are going to be generating high titer vector. So this has really been kind of a breakthrough moment for producer cell line. So yes, and obviously, it's not specific to a potentially specific across the board. So with that, I shall hand back over to Nick.

Great. Thanks,. Dan. That's quite an in-depth summary of our TetraVecta. But I really hope everyone sort of the key take-home messages from this. I mean the first we can produce higher quality, higher titer vector, the only benefit of more potent antiviral vector particles using a TetraVecta. And this really leads into it allows for the successful manufacture of previously nonviable LV based products. So we mean this by, when you look at the commercial aspects of producing viral vector for a therapy. If your titers are very low. If it may not actually be a viable product in the long run. But some of these technologies, some of the enhancers, the reduction of the pricing, maybe the TRiPSystem you can actually significantly increase your prices to make it much more attractive proposition to produce the spectrum and use it to treat patients. The added ability of onboarding large and multiple transgenes is an added benefit. There is a trend in the field now. So just sort of mono CARs, we're looking at dual CARs on [indiscernible] antiviral vectors this is just simply because science is moving on, and there is more need for sort of check price inhibitors and multiple targets that people are looking to address in oncology. And again, the development of high titer stable producer cell lines has been a real issue in the field. It's successful for some products, particularly with lentiviral vectors. But there's always been a problem with the Kymriah gen receptors. Be it due to toxicity or other splicing issues. But using the TetraVecta, the TRiP System, we're now able to develop these [indiscernible] for transient systems. And this really paves the way for really big indications where you're going to need a lot of vectors and very consistent in its production as well. And this consistency really leads in as well to in-vivo applications. Now there is a movement in the field to start exploring the possibilities of in-vivo CAR therapy with the vectors directly injected. Into the patient. The opportunity, et cetera, vector, like Dan has highlighted, the improved safety with a super LTR, the use of the TRiP System to stop any transgene being expressed and incorporated into the vector which may result in off-target. And also that produces cell lines this should greatly improve the quality off vector the consistency produced residual DNA. So I hope you really enjoyed the vector. I have you learned a lot of it, a lot from. There's a lot of particular biology, but this is recorded, so you can always go back and rewatch it or ask us questions in the Q&A. We're also happy to talk to anyone from research or industry that are looking to evaluate TetraVecta of any of these components, and we can be reached of the partnering at oxb.com. So thank you all for listening, and we're happy to take questions.

Well, first, I'd like to say thank you very much, Nick and Dan, for that very insightful presentation. I hope everyone enjoyed it. [Operator Instructions] So while you were speaking, Nick and Dan, we did receive a bunch of questions, so let me start with this first one here. This audience member is asking, do you have an example of a product which has been improved using OXB technologies? That's a good question. Who would like to start with that one?

Yes. So [indiscernible] the question was approved. So obviously, we have technologies being used in, customers are using GMP and those products are being used in trials. So I don't know if I answered that question.

Nick, did you want to add anything to that?

Yes. I mean the TetraVecta has not been in a clinical trial yet. But some of our other components on the U1 technology is being used in GMP and we'll be going into patients shortly. But this is very cutting-edge stuff. So we're keen to talk to anyone that would like to adopt these technologies and et cetera vector and push it forward. We have several partners we're evaluating at the moment we hope push this for very single day.

Let's go to our next question. This audience member is asking how long does stable cell line development take? And what are the advantages other the, sorry, what are the advantages of their transient?

Right. So stable cell lines, the core stable cell line work actually takes about 6 months to generate more cells. There is some work upfront where you will be doing some cleaning and optimization after we've got the stable cell line. So we can expect to have a very good producer cell line within a 12-month period. If you want to look at things like stability of the cell line, kind of important in manufacturing where you'd be working with without any sort of selection antibiotics, that can push it out really to 18 months. But if you have a cell line in 6 months, you can have a fully sort of qualified and stability cell line sort of in 18 months already for GMP. But the [indiscernible] have asked you to it really is once you've made you don't think plasmids anymore. It's it's on demand, really, you can scale it up 1,000 liters, 10,000 meters if you want a 2 inspection process. And you have a better residual profile. I mean the cells are less stressed, doing a big transfection step you have any residual plasmid DNA. So you get a better quality products at the end, and it's more consistent. It's coming from a clonal cell line so you can be assured of the quality of the vector coming out at the end of it.

Let's jump to our next question. This audience member is asking, do you have an example of a derailed product development because of transgenes contaminants? Who would like to tackle that question first.

Yes. So Yes. So we have experience with an issue like that. Obviously, we can't be specific, but we are aware of a client who, in fact, infact harking back to that example that I was talking to about tissue-specific promoter that they didn't believe that they would have a problem with the gene of interest during production. And obviously, that became a problem because of a splicing issue. And so because that gene of interest was just accretive protein, they ended up having to develop another step in a downstream to remove that pricing because they didn't want to be administering it. The other point on delivery. And so there's obviously costs associated with that, the delays associated with that. So we obviously are, we do, we can point to examples why this is a problem.

Okay. Anything to add there, Nick, before the next question.

No, I think Dan covered it perfectly.

All right. All right. Here's the next question. Also a very simple one. Does TetraVecta cost more?

I can take that. Short answer is if you manufacture with us. No. It doesn't cost you any more at all.

Can you repeat that again?

The TetraVecta, if you manufacture with OXB, it's part of our platform offering. So it does'nt cost any more to have the technologies. We are willing to discuss discrete licensing opportunities and if people would like to see that for research or commercial users, please contact our BD and our partnering.

Okay. Perfect. All right. Let me squeeze in two more questions here. I think we can do it. At the TetraVecta system enables the production of more particles with full-length RNA, can you expect higher QPCR titer?

Yes. So that's, I think, sometimes is the answer to that I think it depends a little bit on the problems of the agent interest might be giving you sleep. Depending on what's included, I know there might be different content, the stability might be different. So as I said before, it is possible to generate high titer or high-ish titer when you have significant splicing going on. Was that we have shown examples where we are able to increase the [indiscernible] RNA. So I think it's possible, and I think we're introduced whether or not we shifted the bottleneck. So maybe the bottleneck is generating enough particles. So that's obviously something we're looking in see.

Okay. Let's jump to another question here. To what manufacturing scale have you tested the TetraVecta System?

TetraVecta been up to our 50-liter scale at the moment. We've not taken it to 200. Done a lot of work with a 7-liter scale as well, which is like our validated scale model, it performs very well. It's just it's a viral it's a vector system. So if you can scale it from 7 liters, we can say to 200 liters or more service is tried and tested in different scales.

Okay. Perfect. I'll jump to this question here. Can you share how the TetraVecta System is targeted to specific DNA sequence of the target cells? Very specific.

Do you want to take on that?

Yes. So I said it is a bit confusing there. So we're not saying that tetra or Lenti in general is targeted. It doesn't go to a specific lows in the cell. We, we know that end of our vectors have a particular distribution into active gene. I think the point we were trying to make on that slide is that it's an orchestrated process, but it's well understood. And we know in the field, there's a long history of the safety around that. And obviously, there was a concern about census, but it most like the kind of the numbers of integration of NTB per cell that would be significantly higher policy actually achieve. But what we are saying is that, that process is orchestrated and the payload is protected, unlike perhaps other modes of gene therapy where you effectively end up with a naked piece of DNA in the nucleus, whether that's, I guess that's true for AAV but it might be true if you're trying to use a donor DNA for Crisper or something. Episomal DNA hanging around in cells for a long time. This is what the regulators are now starting to ask around but the propensity of rearrangements and reshuffling in random integration during a subdivision. And what we're saying around Lenti that it's much, much more orchestrated that process. And so the possibilities for the cell or for your cargo to be affected in trade is much, much reduced. And that's why we think it's still important to be talking about lenti in the world of gene in CRISPR. We still think those is relevant. I think on deliver large payloads with gain of function, right?

Okay. Detailed answer that again. Jump in to the next question. This audience member said, I did see transcriptomic studies on a slide. What approach or platform is taken to do that?

Right. This is a question. I'm not a [indiscernible] for that. But we've been doing RNA sequencing. We've generally been using short read, generation sequencing. And I think it's probably mostly on our alumina platform. We've been looking at Nanopore long-read sequencing as well. But as we're still developing that. Going forward. But an interesting question that transcribes we started pairing that with [indiscernible] as well. So looking at the transcript over of the cell and then comparing it with the protein to see how the RNA is comparing with the protein expression in the cell.

Okay, right. squeezing the last question, can any of these technologies be used with other vector types as audience members asking?

And. Yes. Sorry, we have previously showing that trip system can be onboarded into AAV and [indiscernible]. So we anticipate it to be efficacious in pretty much any borrow-based system. So AAV is something that we've, a lot of assets, a lot of the optimization around TRiP and those plug-and-play promoters were actually done in AAV genome because we kind of wanted to make those relevant. So those are all available. We'll essentially supply a promoter UTR TRiP Sequence but it's the same in everything from motor setting. So I think it really kind of standardizes what would be supplying. So and those focuses will be can be made available in AVG and anti genome and whatever.

Okay. All right. I know I said that was my last question, but I'm going to squeeze this one in here. Have you optimized LVs for the coexpression of multiple genes of interest either with 2A peptides or multiple promoters? Can we get a quick answer to that one?

Yes, we have.

And that was the answer.

It will also, obviously, with 2 leverage trip those open reading frames are all linked, but we also have trip running with IRS elements as well, just to sort of TRiP piece about we have experience with all sorts of complex payloads be different promoter than orientation goes on.

Okay. Perfect. Thank you for making a short and sweet. Well, thank you very much for those answers. We have reached the end of the Q&A portion of the webinar. If we couldn't attend to your questions, the team at Oxford BioMedica will follow up with you after this presentation. And if you have any further questions, please go ahead and utilize the e-mail address that is on your screen partnering at oxb.com. So thank you, everyone, for participating in today's webinar. You will be receiving a follow-up e-mail from Xtalks with access to the recorded archive for this event. Additionally, there is a link to view the recording of this event, which you can also share with your colleagues so they may register for the recording here as well. A survey window will be popping up on your screen. Your participation is appreciated as it will help us to improve on our further webinars and get even more out of today's presentation by downloading the supporting materials that we have available for you under the handles tab. There's OXB TetraVecta brochure and then I believe a PDF of this presentation. So please go ahead and check that out. And now please join us in thanking our speakers, Nick and Dan for that very insightful presentation and for answering all your questions. We hope you found this webinar informative. It has been my pleasure to be your webinar moderator on behalf of the team here at Xtalks. We thank you for joining us. I'm Sonia Hunt until next time. Please take care, and bye for now. Thank you, Dan. Thank you, Nick. Bye, everyone. Thank you everyone.