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All right, afternoon, everyone.
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We'll change gear a little bit now, to focus more on the upfront workflows for single cell analysis and for genomics.
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I'm Martin, I'm the CEO of Levitas Bio.
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As a company, we're really focused on the upfront sample preparation.
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So I think all of you are well familiar with the new trend in large data models, very large single cell sequencing works that are ongoing at the Broad, Zuckerberg, and a lot of Pharma companies.
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But fundamentally to be able to generate large data models, we need lots of samples.
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And unfortunately until now, sample preparation has actually become one of the main bottlenecks for being able to generate large models with a large biologically relevant number of cohorts of samples coming into the workflow.
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And we really focus on solving this bottleneck.
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We're kind of ambiguous to what type of sample we use.
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We can use anything from blood, PBMCs all the way to cell and development and dissociated tumour samples.
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And downstream we feed single cell workflows, we feed proteomic workflows, flow cytometry.
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But it's really this idea of dissociation and enrichment in large number of samples in an automated workflow that we focus on.
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And so if you think about sample preparation today, it's a pretty archaic method, right?
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Things have not changed in over 30 years.
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Whether you're using basic dissociation, whether you're using beat-based platforms or flow cytometry, even those methods are pretty archaic.
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It's one sample at a time, lots of hands-on time.
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And the platforms today do not allow you to multiple steps in one go.
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And more importantly than that, depending on the operator skill, depending on the method used, you can actually perturb the sample to a quite extensive amount.
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Your gene expression signals might change, activation states will change.
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And quite often the sample you end up with is actually worse off than what you started off with.
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And so as a company, what we've done is we've automated this step of the workflow to allow you to process hundreds of samples at once, be able to multiplex multiple steps into the workflow and be able to produce data with the highest yields, highest accuracy sensitivity possible.
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And I'll walk you through how some of those applications work and how as a company will be able to deliver this.
2:34
But effectively our key go to market with this with this approach has been to offer you 100 times the sample throughput.
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So for the same amount of time that you would take to process one sample today, you can process up to 100 different tissue samples at once.
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Our all in cost for enrichment selection cell washing is $10 per sample to get an order of magnets less expensive than was in the market today, and on average would generate about 500 times more data from the same sample.
3:05
And our customers over the last four or five years have already validated our workflows.
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So the top 12 to 20 Pharma existing customers, some of the key CROs, and a lot of research hospitals and universities have validated that they can improve their throughput lower than costs.
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But more importantly, with our touch-free prep method, they can maintain viabilities, maintain gene expression and be able to provide biologically relevant results.
3:36
And what do we offer, really across the entire product line, are three basic steps.
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One is viable cell enrichment, allowing you to pull out just a viable fraction of your sample, get rid of any debris or contaminants that are unwanted in the sample and allow you to pick specific cell types if you choose so.
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So for example, if you're only interested in CD45 positive or negative cells, CD34, CD4, we can allow you to include that specific selection of cells within that population.
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Now we also work with nuclei cells.
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If you're interested in organoid workflows and PDX workflows and nuclei cells, all of those samples and workflows are compatible within the platform.
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So it's not just cell-based systems that we offer.
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And so what we have is a complete workflow that starts with tissue dissociation and nuclei extraction.
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We have two platforms.
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There are two sample or eight sample based.
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We can do one, four or 96 sample workflows depending on the modularity and the platform you're trying to use, and the software, effectively the reagents that supports everything is a variety of kits that are specific to your application.
4:49
So whether it's a Nuclei extraction Kit, a LeviSelect Kit that allows you to select for specific immune populations, Residual RBC Depletion Kit or things like Myelin removal Kit, all of these reagents are compatible across all of the platforms and in one step allow you to multiplex all of those steps.
5:08
And so the premier platform that we launched earlier this year, it's a 96 well based system, allows you to process 96 samples at a time in about 45 minutes.
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And it actually operates for two different modalities.
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One is cell enrichment and nuclear enrichment.
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The other one is cell washing.
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So you can actually use it as a full cell washing platform for either CITE-Seq or flow cytometry applications, same workflow, slightly different results.
5:35
And so really the key three applications that we focus on today are your flow cytometry,
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where you are able to wash away antibodies and debris in one step, scRNA-Seq where it's specific debris removal lifestyle enrichment and cell selection for RNA or DNA based application, and then CITE-Seq, which basically overlaps those two methods where you have cell washing and cell enrichment in one go.
5:59
The entire platform is really based on innovation of George Whitesides.
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If you're not familiar with George- prolific Harvard inventor, invented the MRI- and the way the method actually works is based on George's invention of contrast agents.
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So basically how a contrast agent works in the body.
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We're able to create multiple different density gradients across individual wells in a 96 well format.
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So you can almost think about each of these micro wells with a 96 well plate as the analytical centrifuge.
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We're using very specific magnetic gradients.
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We're able to select specific cells of particles of interest.
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So in this case, what you're seeing, the real red dots are the viable cell population that is being selected away from debris dead cells and let's say CD45 positive cells.
6:48
This occurs 96 samples at a time, takes about 20 minutes.
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So the entire workflow from a user perspective is you simply dispense 96 samples at once into a 96 Volt plate, that goes into a robot, takes about 20 minutes for the samples to levitate.
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And you can see the live fraction levitating right there while we're imaging.
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And then post 20 minute levitation, you simply collect only the cells or nuclei of interest ready to go with your downstream workflow.
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So if you work with parse or scale, for example, the output from this tube goes directly into a parse of scale 96 indexing application or indexing plate and you're often running into your bulk coding and RNA.
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If you're using this for PDX models or for organoids, obviously the 96 output plate can go into your specific workflow of choice.
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There's no additional processing required.
7:39
And so I'll give you an example of how significant of an advantage we offer.
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If you're working with neuro samples, brain samples, Myelin is your nemesis.
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Traditional workflows to get rid of Myelin are painful - require multiple centrifugation steps.
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We collapse it into a 2-step process.
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So just to wash and process 24 samples, we're looking at about a tenfold time saving, in terms of time hands on, overall time and steps required.
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If you're more in the oncology market and you're doing things like live cell enrichment, debris removal, CD45 depletion with traditional beads- there's over 50 unique steps.
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We collapse all of those into a single step that's multiplexed and includes all three of these steps into a 20 minute workflow.
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And for 96 samples at a time you're looking at a twofold, or two orders of magnitude decrease in cost, time and improvements in throughputs.
8:39
Now obviously all of that is great if the data doesn't speak for itself.
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Majority of the comparisons we do in-house is to compare to gold standard with our traditional either bead-based or centrifugation-based methods.
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And for this example for Myelin workflows, we're have a roughly tenfold improvement in Myelin removal within the same neuro sample.
9:00
These are mouse brain samples that we process in-house, and again all of this at 96 times the throughput.
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One of our biggest advantages, is our ability to work with very few cells regardless of starting cell number.
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We can go as low as about 5000 cells and surpass over a million cells per sample.
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Whether you're working with the DTC or a PBMC sample or bone marrow sample, regardless of the cell number, the efficiency and the yield of the process are exactly the same.
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So unlike traditional methods, we're not sensitive to cell input.
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So we can go very low or very high and maintain the same linearity.
9:37
And all of this data is generated by one of our partners, Discovery Life Sciences that uses the platform in-house.
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Now obviously one of the other key requirements is that as well we levitate, we want to make sure we're not biassing the sample.
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One of the key requirements to make sure that the population representations maintained.
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And as you can see, regardless of whether you do manual washes or levitation, you're not biassing the present the populations.
10:04
More importantly, as we go into more specific populations, not only are we maintaining the same fraction of lymphocytes in this case, but the percentage of life lymphocytes is significantly higher than it would do with any manual washing or enrichment method, right.
10:20
And obviously, as we kind of keep going into more and more specific applications, these are flow cytometry examples where Tregs are kind of a gold standard for being able to maintain population representation.
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And again, you can see whether we use manual workflows or our fully automated workflow, not only is the percentage of Tregs the same, the actual live fraction of the sample is significantly higher than would be with a manual method.
10:47
And finally, we always joke that the debris removal is free, whether you do live cell enrichment or cell washing for flow cytometry of CITE-Seq, you automatically also remove the debris no matter what.
10:59
So you can see here after manual wash, we got about 40% debris.
11:03
After our wash we should be down to below 10%, right.
11:06
And again, this is on top of your cell enrichment and cell washing that happens automatically.
11:12
Now one of the big adventures as I mentioned before, regardless of cell number, starting cell number, our reels tend to be in the high 80s for PBMC samples, high 60s for DTC samples and those kind of stay the same regardless of input cell numbers.
11:30
And overall, again, all in per sample, our cost is $10.
11:34
And again, we really highlight the cost.
11:37
It's because as people start talking about analysing and processing tens of thousands of samples, the work implications, the cost implications, and the ability to run hundreds of samples per day is going to become paramount.
11:50
And we believe that's going to be the main limiting factor for how quickly these large studies can scale up and start to produce results.
11:59
And so as a final summary, really, we play across multiple applications, not just genomics.
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We're getting more and more into PDX and organoid workflows.
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A lot of our customers are now enriching organoids for viability or size selection.
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We also have a PDX programme would have shown significant improvements in terms of engraftment rates post enrichment.
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But in general the key to the technology is really to enable you to kind of throughput and workflow improvements while maintaining biologically relevant information that currently do not exist in the market.
12:33
All right, any questions?
12:45
For the 96 platform, in each single well, how many cells you can put it there to start with?
12:51
Anywhere from 10,000 to over a million. For PBMCs
13:00
you can go higher.
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For DTCs, about a million.