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Right, thanks for coming along everyone.
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So here Unchained Labs, we are super excited to talk to you today about the Sunny Suite, which is designed to give our customers an end to end particle synthesis solution for LNPs starting at development and moving all the way through into manufacture.
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We're also going to be welcoming a new member to the suite today.
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So I'm really excited to share that news with you.
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So as a quick introduction, Unchained Labs is a company that aims to provide solutions to tough problems in the biotech and biologics industry, and it aims to do this by focusing on innovative products with efficient workflows.
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This is just some of our portfolio and we do everything from protein stability platforms, automated buffer screening and exosome analysis.
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So there's something for everyone there, but today we will be primarily talking about the Sunny Suite and their use in LNPS.
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Our Sunny Suite systems are designed to unleash your LNP synthesis, help you quickly shine a light on your winning formulations and scale them up with ease.
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So say goodbye to single use cartridges and hello to fully automated microfluidics.
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But before we get into the details, we'll have a quick look at the industry as it stands.
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So as you'll hear, you're all fully aware of the success of mRNA LNPs, especially in the COVID vaccines, which has demonstrated their significant wider potential for use in medicines ranging from cancer vaccines through to genetic editing.
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These versatile particles allow the efficient delivery of genetic material into your cells where they can be used to deliver instructions, interrupt processes, or as we said, edit DNA.
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Pretty much any LNP researcher is aiming to end up at some kind of clinical production.
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This will often start off with large scale formulation and material screens to find the best candidates, the optimization of those candidates, and then moving across into GMP manufacture.
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So that's multiple stages, probably involving multiple labs, each with different burdens in terms of the number of samples you need to run, the volume of those samples, and then the regulatory compliance you need to produce clinical material.
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So we'll start off unsurprisingly at the beginning with the formulation screening.
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So lipid nanoparticles are typically composed of four different lipid components, though that's rapidly changing.
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So the ionizable lipids, which is where most of the IP is held in is the most significant component, the structural lipid, the PEG lipid, and cholesterol, those are your typical four.
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And more we're seeing additional targeting, moieties being added to the system as well.
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And all of those components have to be the right components in the right amounts to ensure the correct particle performance.
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And that's everything from how stable they are in storage through to how stable they are in vivo, the immunogenic response they generate, their cell uptake and then their endosomal escape.
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So there's loads to think about there.
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And that's just increasing as people find new ways to target these to specific cells and organ types in the body.
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And that's a huge challenge.
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The complexity of the formulation stretches far beyond just these 4 lipids.
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There's also the cargo, which is a huge field of research in its own right.
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And then there's all the processing parameters which can affect the structure and efficacy of the final particle.
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So that's the mixer type of the flow rates you are using, the NP ratios and then of course the excipients and the storage of the particle.
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And what's our solution to this?
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That is the Sunscreen.
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So Sunscreen runs off 96 well plates for your radiation inputs and your sample collection.
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So it can run a fully automated set of 96 experiments in under 6 hours.
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So that's all with microfluidic precision.
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The system uses driving fluids, so typically ethanol and a buffer to push your sample slugs through the system.
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So that means there's no dead volume in the system.
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So you can really keep those reagent volumes low.
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We can say you can go as low as 100 microlitres per input, so as little as 200 microlitres per experiment, typically 400 microlitres if you're working at that three to one ratio of mixing, which we find most of our customers are.
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The microfluidics are fully reusable, and the method is designed to be scalable in between the different pieces of equipment in our range.
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Precision manufacturer of LNPs needs precision mixing.
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So the mixer that we provide with the kit we call our Sunnies and these are glass microfluidic mixing chips.
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They come in a range of different mixing types, so you can use whichever solution suits your production the best.
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We went with glass chips because this comes with a host of benefits.
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They're highly chemically compatible, so you can use a wide range of materials and solvents with them without worrying about damage to the chip or to your reagents.
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They're also incredibly smooth.
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The surface roughness of these chips is approximately 5 nanometres, and this is compared to with about one to three microns with an injection moulded chip.
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So this means that there's much less material build up over time in the chip and at the mixing junction.
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And it also means that the cleaning of the chip is really efficient.
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And all of our pieces of equipment are designed to have automated cleaning cycles in between every experiment to make sure that channel is clear after each experiment.
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So let's see if this works.
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There we go.
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Perfect.
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So this is an example.
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We ran on the Sunscreen with two dyes, a blue dye and a green dye.
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And you can see them being pushed through the system by the driving fluid meeting on the chip mixing and then being pushed out by driving fluids, which then helps clean the system.
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And we've tested all the volumes that we push through to the system to ensure that's enough to displace any RNA that's left in the system after the mixing event.
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Sunny Suite comes with simple to use software, so there's just an Excel spreadsheet that you use to input your experimental variables, and you can edit that then in the lab, in the office, or even at home to lay out all your experiments.
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You then just upload the spreadsheet into the software and then it will automatically run all of your experiments sequentially.
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When it's doing that, you can just walk away and let it do its job.
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Of course, if you're running this many experiments, you need to be really sure that any variation you're seeing from formulation to formulation, whether that's in biophysical characteristics or in efficacy, is down to the formulation itself and not the process you're using to make it.
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So to test this and demonstrate this, we ran a plate of 96 different formulations, six of which we intentionally removed the PEG from to induce catastrophic aggregation.
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So you can see those in red, and you can see the samples after that have returned down to that baseline size around 90 nanometres and PDI around 0.14 with no trouble.
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So you can really trust that our cleaning procedure will give you really high experimental fidelity from well to well.
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The CV for these experiments, the coefficient of variation was 1% between all of the formulations labelled in green.
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So you can really be sure that when you're switching from formulation to formulation, if you see differences, they are down to the formulation and not down to the equipment.
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And this means you can carry out large scale screens with confidence that what you're seeing is what you need to be seeing.
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So here we did 32 different formulations in triplicate.
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We varied the cationic lipids, the structural lipids, the pegylated lipids, identity and amount.
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So that's those characteristic down slopes in particle size.
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You can see across each formulation group as we increase the peg lipid content.
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And this lets you pull out really interesting data really rapidly.
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So this is the effect of structural lipids on your PDI.
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We found that DSPC typically gave lower PDIs than DOPE and that DOTAP typically produce smaller particles than DDAB.
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In this case, we stopped here, but obviously you'd be probably taking that through to in vitro or in vivo screening from this point.
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And you might be thinking that is a lot of analysis and it definitely was.
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I did all that in the lab in one by one DLS because that was before we were acquired by Unchained Labs.
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But Unchained Labs provides the excellent solution to that which is the Stunner.
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The Stunner is a 96 well plate based DLS and UV vis system.
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It uses just two micro litres of per sample and then that gives you information on the particle size, the RNA concentration through UV vis and an algorithm that extracts the signal from your RNA from the lipidic signals.
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It can also give particle concentration because it uses multiple angle DLS.
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You can get accurate reads on that and then it can calculate average number of RNA particles, RNA per LNP.
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So that's a really unique feature the Stunner has.
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It does that all pretty rapidly too.
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And then once you've screened your particles, of course, as we said, you can move those into in vitro or in vivo testing.
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So this is some work primarily done by our customers, both on a luciferase assay in cells and then the same particles put in vivo.
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And so the perfect particle, your application is out there amongst all those formulations and Sunscreen is designed to help you find that as rapidly and as cheaply as possible.
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And so what do you do once you find your leading candidates?
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Sunshine is the ultimate in rapid semi-automated formulation screening.
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You just load your samples into the sample loops at the bottom of the kit there and then the equipment will aliquot that into up to 10 experiments.
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It'll run all of those experiments automatically.
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So whether you're looking at what the effect of different flow rates, the effect of changing your dilution level or the effect of changing your flow rate ratios, do all of that within 15 minutes.
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And then it has the same automated screening, automated cleaning cycles that Sunscreen has, and it will run samples between one mil to fully continuous manufacture.
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It uses the same reusable microfluidics.
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And it's designed for really easy transfer of protocols from Sunscreen to Sunshine.
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And so once you've run a set of experiments, you just load more samples into your sample loops and you get going again.
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As I was saying, it uses the same microfluidics, it also uses the same software.
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So there's really no learning curve in between using the Sunscreen and using the Sunshine, and you can just plug in those parameters that you used in the one piece of kit and plug them into the other and transfer that method straight across.
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Using Sunshine, you can focus in on the particle properties you really care about so particle size, PDI loading efficiency, and of course efficacy in vivo and in vitro.
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Because they're so quick to run, you'll stay incredibly rapidly through process development to the need to scale up.
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And this is just a set of experiments we did with some DOTAP based LNPs looking at the effect of different flow rate in triplicate.
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So once we'd settled on the conditions we wanted to use using that flow rate screen, we selected 9 millimetre because that produced very nice 80 nanometre particles with low PDIs and high encapsulation efficiencies.
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We then decided to take that up to continuous manufacture.
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So here you can see we ran a batch of 45 mil which we separated into 1.5 mil aliquots.
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You can see the consistency of sample size across that run.
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And we are now happy to introduce to the family the Sunbather.
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So the Sunbather is our solution to the GMP manufacture of LNPs.
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And again, the idea is you can take a process straight from Sunshine running in continuous mode, run that in Sunbather with no additional process validation or changing of technology necessary.
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So Sunbather that runs up to 1.8 litres per hour.
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It's got a fully replaceable flow path with extractables data.
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You can use the same range of mixer types.
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It's got 21 CFR Part 11 software, of course, and it's designed to make your life very easy when you're moving into GMP.
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And that is because as I've said, at every stage, you're using the same pumping technology, the same mixing technology, and all of the top software to make your life incredibly easy.
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So you can trust the process from 800 microliters all the way through to 8 million.
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So come see us at booth 10 if any of that has interested you.
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We can talk about LNPs, and we can talk about any of our other solution platforms.
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We are just in the corner near the inflatable Flamingo.
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If you're looking for us, I'd be happy to take any questions.
