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Thank you.
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So thanks for attending this presentation from Refeyn
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I'm obviously not Salma and my name is Tobias.
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I'm a technical sales specialist from the South of Germany, Austria and Switzerland.
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And today I'm showing you some news about mass photometry.
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So in this presentation, we'll start with how to accelerate your analytic from antibody measurement, also how to accelerate your projects in the development, but also how to derisk your project in terms of discovering things going on in the early stage of your research.
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So Refeyn is a very young company.
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It was founded 2018 as a spin-off of the University of Oxford.
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And now we are already 200 people working all over the world.
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Main development and research is still in Oxford and today I’ll tell you why the best way is to stick with mass photometry.
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So it's an optical measurement, so we don't have anything to do with columns and also site is more like having issues with clocking columns.
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But also we don't really have problems with skewed UV absorptions in a way columns interaction might happen during the purification.
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But there is no column, so there are no column interactions at all.
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And also we have a really low sample demand.
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We just need nanograms of your sample to get all these awesome results.
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So who has heard of mass photometry before or Refeyn?
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Great.
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Mass photometry is a single molecule method.
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We don't need anything to mark our molecule.
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So there are no markers involved.
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So also no extra working step.
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That's the good thing.
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And we have a really low sample consumption nanograms and we just need some micro litres for nano gramme molar concentrated sample to measure it.
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The way mass photometry works is so biomolecules are charged by its nature and also glass surface has a certain charge.
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What is going on in that case is we have our biomolecules in solution, they start to interact and also bind to the glass surface and when they bind to the glass surface that is the first step in the analysis, but it's done automatically.
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Why it's called mass photometry.
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So we have an optical unit where a 488 nanometre laser is placed and this laser is illuminating the biomolecule or the drop where your biomolecule is in.
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And most of the light is just getting through the drop.
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Some of the light is getting reflected.
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But every time when one of the biomolecules is binding to the glass surface, you get a scattering signal.
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And this scattering signal is displayed here as a so-called point spread function.
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This point spread function is directly proportional to the size of the biomolecule.
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And the great thing is it's that the shape of the molecule or what the bio molecule contains doesn't affect that.
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So if it's round long, if it's contains DNA, different products, doesn't matter, it's just the size and the mass of the biomolecule and that's what you see and what you get as an output.
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So you just add the sample to your measurement surface, and you get mass histograms.
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So in this case we have three species and that's the overall counts for each mass in solution.
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So the way it works is really simple.
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So one of our CPOs said once that everybody who's able to operate the pipette and can use a nano drop is able to use a mass photometer.
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So people like me can do it too.
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So that's pretty good.
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And can we click, is the video working maybe can you click on the video please?
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Yeah, thank you.
6:05
So that is how the measurement is done.
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You open the lid, add your sample to your cover slip and then you mix it really well that you really have a homogeneous sample.
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And after that the just close the lid, press the start button and immediately when you press the start button, all this binding events are coming up.
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You measure for one minute and after this one minute you can do the analysis of your sample.
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So there is not really much waiting time.
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You can do it, get everything straight away.
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And what you can find out here is in terms of antibodies, you can for example see really well if your antibody is in the monomeric state or if it's in the dimeric, trimeric, tetrameric state.
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And that is something you get straight away.
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And what we want to avoid in this case that high molecular weight side products during the antibody production and development are somehow getting direct on in the process and we just need some nanograms of sample.
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So even in the early stage we can discover if something is going on here.
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For example, we have our example of a size exclusion.
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The size exclusion it's more like the best thing you can use to purify proteins and biomolecules, but you're always unlimited to a certain molecular weight where you have the best resolution.
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That is something we don't have.
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We have a linear resolution from 30Kilo Dalton up to 5 million kilo Dalton.
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So we avoiding that and we just need nanograms to analyse it.
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So even if you have more like the size exclusion and you look at the different fractions, you can just measure all the little the fractions again with the mass photometry to see what is in there.
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So what does this tailoring this little peaks or right in front, what does it mean for you, your experiments and your project?
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So everything what is somehow indicated or caused by a shift of a mass, you can see the product.
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You can see also here some aggregation going on, if it's forming a dimer, but also if for example, there is a misassembly or also something missing.
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That is everything what you see while performing mass photometry.
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What is also really nice.
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I love this example.
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It was done together with our industry partner.
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So they measured 14 different proteins.
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They made a stress test with light and temperature and peroxide.
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They measured in total 169 times.
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All the measurements were done in half a day and they used 7.5 nanogram sample with size exclusion you would spend much more time.
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You can automate it, yes, but you need also more sample for sure, but it's more like an orthogonal method to see what's going on in your project.
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What we have here.
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It's also when you somehow start what you see in the mass photometry like you also see really early what is different collection, what your sample is doing when you do this clone screening, what is the best clone, which clone is for example producing more side products and other ones.
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That is something you can't cover or discover with this SEC unfortunately, but over all the other techniques available on the market too and every technique is really well known on the market and had has to be there.
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We have here more like some 3 prominent single molecule methods.
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First the mass photometry, second the AUC and the DLS.
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What is striking here is with the DLS the monomer has its issues that it's not resolved.
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With the AUC you get a pretty good similar result, but you would have more like the reason you're losing the resolution with the higher aggregates.
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But the most striking is the sample consumption and also the time consumption.
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If you work in the lab, you have your cup full of your antibody.
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You want to see straight away what's going on there and not wait hours, days till you have a result.
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So even in the early stage you can see why that is happening and also for example, do some buffer optimization or do changes to the protocol to see what's going on.
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Also accelerating project timelines, so if you see really early why something is happening or also why it's behaving like this, you can stop a project much earlier because you have this information earlier.
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And instead of just dragging on the project longer and longer, at the end you find out what's not worth it. In terms of ADCs, what we can see here as well.
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So that is a little more tricky.
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You have the single antibody and then you add to conjugate and in a way you would see if you have everything in the same solution in the same sample, you might see a small little shift.
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But here when you have it side by side, you can somehow see OK, the derivate is binding, so there is a shift going on.
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If several of it are binding, it's even better because that mean that you have a clear result.
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And also if the derivate is a little bit bigger, you can also get an idea about the stoichiometry of the binding.
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That is one of the things I really like too, because it shows you how well your antibody is produced in your cell line.
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That is, you can see just looking over here that is the untransfected cell and you don't really need any purification steps, a slightly small purification step in terms of that you have to clarify your sample, but then you can measure the supernatant straight away and see how well the antibody is expressed in solution and that is what it looks like.
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So we have our TwoMP in that case we provide a semi-automated solution.
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You have really great group of software engineers.
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We are permanently working of the improvement of the software and also we provide the consumables as well.
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So the great thing is in this context we have also the software called StreamlineMP that is a software which enables you to analyse high molecular aggregates in the antibody development straight away.
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But from Refeyn we have also other products as well.
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So today it was more about the two MP.
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So it's more like broad molecular range from 30 Kilo Dalton up to 5 million Dalton.
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So you have more like a broad range of applications for that.
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But we also have our SamuxMP and the SamuxMP is a dedicated instrument for gene cell therapy analysis of AAV.
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Well, you can just measure with the exactly same principle the filling of the AAV capsid if it's full, partial or empty as well.
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And the latest product we brought on the market is our KaritroMP.
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It's so-called macro mass photometer.
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It's a little bit different because it doesn't have one focus on the sample, it has several focuses.
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So it's scanning through the sample.
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And the advantage is you can even capture bigger molecules like viruses, adenovirus or other ones.
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But you get also the information about the loading of the virus as well.
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That is more like what you can see here, 40 to 150 nanometers.
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So for them, for that we are as a company based still in Oxford where we have our headquarters.
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We also have headquarters now in the US in Boston (Waltham), but also smaller ones in Shanghai, Kobe and Singapore and also Berlin.
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So we are around 200 employees and the great thing is every day we get more publications or not we our customers, sorry more publications are available.
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And if you look at our homepage you can just search for any application and publication you want, just sorted by the drop down menu while for your application and so see what if there is already something published you're interested in and it's getting I think this year, I thought the last information was we hit the 600 publication mark.
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Now I can see here it's the 1000.
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So it's good.
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And why should you choose the antibody aggregation for your lab.
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It's you can see really early what's going on in your process.
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Are there some aggregates or some other things going on?
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And you can do the stabilisation of your antibodies in terms of buffer screening.
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But also it's a column free method, so you can perform it straight away as I showed before.
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And also the data analysis, really, that's what I didn't mention before.
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In a way, really intuitive.
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What's done by our software engineers.
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The software is permanently improved and really easy to use.
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It's really awesome.
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And also it's just a bench top instrument.
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So you don't really need any extra things around it.
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So not the extra room in a way, you can just put it on the bench top and that's it.
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And also the cost per sample are really low.
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It's good.
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Basically just a glass cover slip with the silicone cassette on top.
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That's what you need to measure your sample.
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So we are today at booth 41.
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Feel free to stop by to get more information.
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I'm accompanied by my colleague Roland.
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He is an application scientist.
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So we are happy to get your questions.
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And yeah, thanks you for attending my talk today.
