Latest / BioSpace / From sequence to scale: Gene editing’s new era in biologics manufacturing
Transcript
- BioSpace Marketing: Hi, my name is Jennifer Smith Parker, Director of Insights at Biospace, and you're listening to Denatured. In this episode, you'll hear from Jack Crawford, CEO of Dimitro, and Magnus Gustafson, Chief Commercial Officer at North X Biologics. We unpack the evolution of cell line development, chhow cells, targeted integration, transpostases, and the collaboration models speeding biologics from sequence to GMP. Jack and Magnus, thank you so much for joining this episode of Denatured. If you can introduce yourselves, that would be great. Hi, I'm Jack Crawford. I'm the CEO of Demetra. I've been running startup gene editing technology companies for about the last 17 years. Demetra is a gene editing platforms technology company, ⁓ and we're focusing on engineering chok cells for cell line development and Biomanufacturing. Thank you so much, Jack. Magnus? And my name is Magnus Kustafsson. I'm the commercial officer for NorthX Biologics. That's a CDO based in Sweden. And I have a scientific background, I would still argue. I started out my career developing monoclonal antibodies and other proteins. I've moved into business development and now work at Northx Biologics with those services. Perfect. Well, thank you so much, gentlemen. So let's start at the very basics here for our audience who may not be as familiar with some of the technical language. So what is cell line development and why is it such a critical step in biologics manufacturing? Thank you. So before we get into cell line development, I I thought I would talk about the step before. This is the discovery. So there's a bunch of types of drug modalities, but ⁓ what we're talking about is biologics. And so the first step is actually discovering the sequence of the protein that the client or whoever's developing the program believes is going to be a good candidate as a drug. And there's various ways to do that. But once you have this is a DNA sequence. Once you have a that DNA sequence, which it could just be a heavy and light chain. Sequence for a monoclonal antibody, then you need to make them protein. And so the first step of that is to make a small amount so you can screen it in cell assays or animal studies. And that can be done using faster methods like transient expression in cells or even newer cell-free methods. But eventually, once you've narrowed down a couple of candidates, then you need to make more material. And eventually this needs to be extremely stable for the production long term. And that's when you really need to get into what people call cell line development or stable cell line development. This is where you take a cell host and integrate that gene sequence. So that it can be expressed on a high level and stably. And so that way you can scale it up and purify that material and then even go into the clinic and commercial. So that that's cell line development in a nutshell. Good explanation for it there. ⁓ Megus, wanted to add anything to that? Yeah, well, I I think it's important to remember that cell lobment is probably the most important decision one can make in the entire product life cycle. That's big words of course, but what you what you start with is what you will be using for the lifetime of this drug. And some of these drugs have been on the market for forty years. So just imagine if you from the beginning had a better performing cell line, how much cost that would be relating to in the end of those forty years. It's so important to get it right. ⁓ you can change ⁓ your your process, you can change buffer or even add it adding purification steps in the process with slight changes though. But the cell line you will not be able to change, then it becomes a new product. It's a re i irreversible decision. I think that's important to point out. Yeah, I mean as you're describing it, this is as you're saying, crucial to your consecutive steps. So you really have to get this right the first time. Jack, you had mentioned CHO or Cho cells, if we can define them actually from the start of what that is. And then why have they been the workhorse of biologic manufacturing for decades? What has made them the industry standard? I think both of us will be able to answer in different ways. ⁓ so Cho cells that stands for Chinese hamster ovary ⁓ cells, and they they were isolated actually back in the late 1950s. And ⁓ so that's one of the strengths of the cell line actually, how long it's been around. And ⁓ it was one of the first cell lines that was used to manufacture a biologic drug. And therefore it has 40 plus years of regulatory understanding and acceptance. So that's one of the the key points. And from a genetic engineering standpoint. There's a lot of advantages of it. It's ⁓ easy to transfect, which means it's easy to genetically manipulate. and ⁓ the genome has been sequenced, so it's well understood. So there's a lot of advantages from our end in terms of the cell line development. But I'm also interested to hear what what Magnus has to say about this question. And from a manufacturing perspective, I I think show cells are not by any means perfect, but they're good enough at everything is kind of why they are so widely used. And I think it's basically five advantages to keep in mind. And those have led to another two advantages. I I will try to explain what I mean. The the first advantage is that shows have a very human like post-translation modification. That's the glycillation pattern. So the human body will see them as human friendly och human like. Det har blid lång frown för a very long times, så därfor there is a a gut regulator acceptance. You know it's you have a smooth path through regulatory using shows and and from the manufacturing point of view scalability of course should be mentioned you you can run this processes in very small scale but you can also scale it to large skales like 20,000 liters. And they grow well in suspension in serum free adaptation and so on. Så it's easy to handle them they're very robust. It's not like very sensitive primary cells that die on you when you when you use them. And the safety profile maybe is the most important point to point out ⁓ they are not so prone to to ⁓ to culture human pathogens so human virus human virus of kors propagate very well in human cells but not så well in show cells. So these five advantages has probably led to that now we have an ecosystem of of show cell manufacturing. There's a huge knowledge base. They established media feeds, analytics and process platforms, equipment providers, and so on. And that's so it's very easy to be one piece of the puzzle in this rather than you know compary. And then the last one is of course that all of these advantages have led to this extremely long track record. The first show-based prote is was activase and it was ⁓ approved in nineteen eighty seven. It was a DHFR negative strain, ⁓ with MTX amplification. And it's still on the market. It's impressive. Yeah, it's a very long track record. And now we have fifteen hundred monoclonal antibodies or so in clinical trials. It's huge. I am curious when you're starting off saying that they're not perfect because everything that you're saying sounds pretty perfect. ⁓ so I just Just to add on that, where where's the fallibility here for show cells? So it's they they are human like in glycosylation, but it's not human glycosylation. So of course it would be better with a human cell line. But in total show cells are probably the best. Yeah. And our mission is to use gene editing technology to improve Chow cells even further. And there there are ways that we can engineer target genes within the Cho that can manipulate the glycosylation, for example. Understood. So just to continue on that, Jack. When you're talking about these gene editing methods, what are the main methods that are used to introduce the gene of interest into host cells? And how do approaches like random integration and targeted gene editing differ? Yeah, that's a great question. It's actually extremely important for the entire program. So the traditional way of getting the gene sequence to stably integrate into the Cho genome. Is random integration, as you mentioned. And so this is basically you have a s usually a circular piece of DNA, and you introduce that into the cells by transfection, and it randomly jumps in in bits and pieces into the genome. And as you can imagine, that's an extremely low efficiency process, which requires the screening of thousands of individual cell clones to confirm that they might have your gene of interest and are expressing the gene of interest. So that takes a very long time and it's a very laborious process. Then the other problem that random integration has is that it's notoriously instable. So then you can't really plan for stability if you're using random integration. So basically what you have to do are these stability studies, which we do anyway, and they're required, but you can't do really anything until that's done. And if you don't have stability, then you gotta go back to the beginning. So that's that's random integration. And that's why the industry is moving away from it. And then you've got targeted integration, which has different flavors. The most common way to do targeted integration. Is with ⁓ what people call landing pads for recombinations and integrases, where you'll get the integration at the same exact site every time, ⁓ which is nice because it's precise and you can kind of predict it. It tends to be more stable. The problem is that the process is very low efficiency as well. So if you have a pool of cells, you can expect under 10% to have your. precise ⁓ targeted integration. So that also takes a long time. So what we use and what I think a lot of ⁓ cell line developers are are going to is ⁓ a mix of both a little bit. The and these are transposases. So we've developed our own transposase and it takes the gene of interest and integrates it multiple copies of the gene. Like you would get with random integration, but it tends to prefer highly expressed and stably expressed sites. So you get higher productivity, higher titer, and then you also get better stability. So that is our preferred method to actually integrate the gene of interest using CRISPR-based technology, which we have our own version called Cast Clover. Is for us more of a cell line engineering ⁓ project and not a cell line development ⁓ for stable integration. So, just to put it in perspective, random integration is like playing genetic roulette. This is done by electoration and you open up the cells, you get the genene, but it never lands where you want it. So then you have to screen for the best clones. I think what Demetra is doing, and other modern cell line development systems. They're using these transposons which gives you predictable high tide to unstable booms and clones from the beginning. That's the beauty of it. Got it. Thank you for laying out the landscape and the different methods of of going about that. So it's interesting to hear, Jack, about the mix of that you're using the transposers for the other option as well, trying to come to essentially a happy medium there. When we're talking about this landscape where companies tend to fall into one of two camps where you're Targeting selling development and GMP as sequential, disconnected decisions, or handing both to a single C DMO that does everything. What are the trade-offs of each? And this is always an ongoing discussion, whether or not you have separate systems or different providers, or you keep it underneath one roof. So tell me a bit about the Demitzha NorthX structure. So if I thought and then you helped me out here, Jack, when I when I che, but I think in the in the sequential of this connected setup, of course the benefit is that then you can select the best supplier for whatever you want. ⁓ so it's very flexible and there's no vendor lock in. But the disadvantage of course would be ⁓ tech transfers and product management, timeline gaps, finger pointing when something fails, that kind of question. And also you might end up with cell line that is not really optimized for a GMT process. ⁓ if you if you go for a single CDMO, meaning cell line development and manufacturing under the same roof, of course, then you just you don't have those disadvantages with handovers and and and so on and maybe faster timelines as well. The disadvantage is that you normally are locked in. ⁓ And it's not so sure that you actually get the bästa provider för både the cellulin and then the latter manufacturing component. So in i vår setup or the Demetra Northsmod, which we are talking about, is that then you get a specialist for both parts. Så Demetra is a trucialist in sällen development and And Northex is has his strengths in in GP manufacturing and we don't try to be best on other things. So I think maybe that's a good advantage. And with this integrated setup that we are working on now, we work as a one company but with two different expertise. Yeah. I think that's definitely true, Magnus. So it's unlikely that a C DMO actually doesn't exist out there right now. Would have all the gene editing tools that we have at Demetra to only improve the chose cell host for future projects, but also for the core cell line development. So Demetra does not need to focus on the regulatory and the types of things that you need to worry about when you're doing GMP manufacturing. Instead, what we get to do is we get to Engineer the cell, make the best producing cell for the long term, and continually evolve those platform cell lines. And what I really like about the partnership we have with Northx is the ⁓ thoughtful way that we can transfer what we're doing at Demetra to North X. Because we have these tools and we know the cell line is going to be stable maybe six to eight weeks from the beginning, and they can do different things, downstream processing and analytical development while we're single-cell cloning. So you can compress the timeline and you can give the customer flexibility on what milestones they want to hit. For what they need to do for their next step. Maybe they need to raise more money. Maybe they need to get it in the clinic as fast as possible. Both of you, I think, really make interesting points instead of of the collaboration, such as yourselves, of the time saved. And I've heard again and again, and especially in my recent white paper on C DMOs, is that in today's world, saving time is means saving money. And that is just so crucial to a sponsor. Yeah, absolutely. And one thing ⁓ that you don't really think about or talk about doesn't get talked about enough, I think, is the amount of paperwork and ⁓ contracts and sometimes licenses, technology licenses that need to be negotiated usually. So if you're a startup and you need to get to the next stage, one month is probably a couple million dollars. And y just to to to put it in perspective and the why I think this is important if you remember I I mentioned this first Chow protein being expressed at that time a reasonable good height it was 50 milligrams per liter. Now with Dimetras cutting edge cast clover curated show cell lines we achieve 18 grams per liter that's 300 times more material from the same Right. And the three hundred times difference in bioreactor is an a huge difference in cost. Absolutely. So this is where the market has taken us in in these forty years. And imagine when we can do this for gene therapies and cell therapies and all these other new modalities. It then they will be affordable. Yeah. Well, gentlemen, on that end we're gonna have to wrap it up. We can talk about this for the next hour. Thank you so much, Jack and Mangus, for chatting with me about gene editing, cell line development, show cells, the manufacturing. And if you'd like to listen to more episodes of Denatured, please turn to biospace dot com. Thank you very much.