0:50
So yeah, thank you very much for the introduction.
0:53
Hello, everyone.
0:54
Suzy and I are going to talk to you all today about our approaches at Thermo Fisher Scientific to improve cell therapy processes with a focus on efficiency, robustness, and preventing contamination.
1:08
You'll have to forgive me if I turn around and cough at some points.
1:10
I have a lingering cough, but hopefully it won't break things up too much.
1:16
I just thought it'd probably be worth us going over a very quick agenda.
1:20
We can probably take the first one off now, right?
1:21
Because we're on that slide.
1:23
But we're going to be mostly talking around focusing on the developments that we've got internally for different products and different processes, hopefully where we can try and collaborate if this sort of space is interesting to you.
1:34
We're really focusing on trying to plug the gap between PD and clinic and then hopefully commercial down the line as well for some nice new bits and bobs.
1:44
And really we're going to be looking at, at least on my side, reducing the risk from media design and then move on onto the QC side of things that just fit across the entire portfolio.
1:54
And then hopefully we'll have a bit of time for some questions, if there are any.
1:57
If not, we can all go get a coffee before the next talk.
1:59
We will see how we get on.
2:03
So Thermo Fisher Scientific are very arrogant and like to think everyone knows who we are.
2:09
But if you haven't heard of us, we are a large organisation.
2:13
I mean, the, the number one here is already outdated, right?
2:15
It, it says 120,000 people, I think what we have 140,000 -150,000 or somewhere around there.
2:20
OK, right.
2:21
So Dinesh is nodding.
2:22
So yes, we have quite a few of us.
2:27
We have a lot of capacity and developments beyond what we're going to be talking about today.
2:32
So this is kind of like a quick whistle-stop tour through some of the bits for this modality in particular.
2:40
But I guess the point of this slide at its core is that we are really trying to leverage the different technical capabilities we have across the entire business in lots of different units.
2:53
And one of the things that Suzy and I do as part of our job is try and pull in those different people that have either been there done that or have a thought process that aligns with ours. Or they may be working on something and trying to navigate something that's a bit more bespoke with the different potential collaborations or evaluations and then feedback into our product development.
3:10
So I think that's exactly the point of this slide.
3:14
I'm not going to waffle anymore.
3:15
We've only got 20 minutes, right?
3:18
And like I said, we're going to be focusing on cell therapy.
3:21
And just on a high level really what we're trying to go about today is about how we're going to future proof ourselves for autologous and also allogeneic.
3:31
So that kind of nicely takes us over to the process side.
3:34
And I think it makes the most sense for us to start with media.
3:39
So today, at least on this slide, I'm going to be talking about some of the different T cell media that we have at Thermo Fisher.
3:48
So the T cell media that we have comes under the family name of OpTmizer.
3:53
There are three different OpTmizer medias, right?
3:55
But the one that I've got here is the newest we call OpTmizer 1, 1 because it's one part.
3:59
It is easy to use.
4:02
And then compared to the previous iterations in that family, this has really been started from scratch using omics and some really neat novel ideas.
4:11
And in order to push the different bits that we're looking at for the development of the media.
4:19
I guess just one note, right?
4:20
We call it OpTmizer one.
4:21
Things like the supplement that used to be separate with proteins that are frozen down, that's all in there, that's heat stable glutamine.
4:28
We made the decision to keep that separate because of things that we've seen such as Glutamax, it's quite common in the HEK and CHO and all those other different workflows, they tend to not always be able to metabolise by different human cells.
4:42
So for us, we prefer to not use that and stick with glutamine and then we keep the glutamine separate because in our experience, we prefer to have consistency across the board.
4:52
You had glutamine at the point of use and then we're not looking at things like age that could come into effect and then cytokines, of course that's going to be process specific as well.
5:01
So OpTmizer one, one part apart from the few things that you need that are specific to you.
5:08
But then I guess in terms of robustness and reducing contamination risk here, this is our first true animal origin free cell therapy media.
5:17
So we've got recombinant proteins in here.
5:20
So better from a regulatory point of view as well.
5:24
And this whole new novel media that's not based off of anything else in the family always mean that we've essentially got a better performance, right?
5:34
So we've got a better performance through keeping early, you know, a better early memory phenotypes in cell culture compared to other media in the market.
5:44
Then because we've got the better cell health, we tend to get a better CD4 CD8 ratio.
5:48
And when we've done some internal studies in our labs, we tend to get better efficiency of the products that we've been working with.
5:59
And then I guess as silly as it sounds, we've also put our medium bags, but we've put that as catalogue options.
6:07
That's not always been the case in the past.
6:09
So there's options for bottles and bags and that was really just to make sure everything's ready to go from the get go.
6:15
So it's a closed system start to finish and be suitable from autologous to allogeneic with the different volumes that we'd need.
6:26
And these bags have got both C flex and PVC tubing on there, so you have options for aseptic welding to keep them as a closed system with any of the different third-party welders you may or may not have.
6:39
Yeah, we thought we would just pull up a couple of pictures to do with that as well.
6:42
So as you can see, we've got a few different sizes.
6:44
We've got a 1 metre on the right hand side and a 10 metre I believe it's 10, could be 5.
6:48
I probably should have looked before the talk, that would have helped, right.
6:52
But we have a few different sizes and there's more than just this as well.
6:56
And these medias and by extension some of the different supplements that will come in bags that have been designed in flexible ways based off of either decanting these into different bags for different systems.
7:11
Because some processes, T regs might need much smaller volumes.
7:16
Yeah, write down 510 mil, but then other people may need volumes that that that are larger.
7:25
So we just try to keep options open.
7:28
And then our equipment which we'll come on to in the next slide, these different medias, the solutions all integrate very easily into that as well.
7:38
And I guess one last point, these all have a one year shelf life.
7:45
So that moves us quite nicely onto the equipment side of things.
7:50
Time's quite precious.
7:51
I don't actually know how we're doing for time.
7:53
Oh, there we are, maybe about 7 minutes in so I need to speed up.
7:55
Sorry, Suzy.
7:58
On the equipment side, I thought we'd just focus on probably the simplest to explain in just a couple of minutes, which would be the DynaCellect.
8:05
We have far more that fits into the cell therapy portfolio.
8:09
It'll make more sense in a moment or so when I come on to that.
8:11
But we're really just looking and with this example, try to explain how we can future proof ourselves rather than scaling out to have consistency, robustness and the speed of processing while trying to reduce the risk of contamination.
8:27
So this slide is a bit of a high level one, I'll be quite quick with this.
8:32
But from our perspective, the biggest challenge in cell therapy manufacturing is the actual manufacturing itself in the supply chain.
8:39
So we have no room for errors, no room for repeats.
8:43
Like I just said, the supply chain is a challenge, so we're looking at trying to close the system, but also make things that are easy to move about and fit in workflows, move between rooms if there's a need for something that goes down.
8:58
And yeah, I guess that's the thought process, right?
9:02
It's flexibility and the DynaCellect tackles this quite well.
9:07
It's pretty flexible.
9:08
So this is a picture of it.
9:11
It's also on a stand.
9:12
We're on booth 10, by the way, you've probably seen us with a big gigantic piece of equipment and we are trying to be modular, right?
9:22
So some others in this field have fantastic ideas, right?
9:26
You have one piece of equipment, you press go and, and, and it runs start to finish.
9:29
I, I think it's brilliant, right?
9:30
It's hands off.
9:32
Our approach is to try and place things so that it could be flexible.
9:37
So if you needed to pivot inside your business, move autologous to allogeneic or allogenic to autologous, there are options there with the same pieces of equipment, but then also as you scale up with different volumes as well.
9:49
And then I more importantly probably should have started with it, is that we have single piece of equipment for each step so that when that step has been completed, that equipment is free up to repeat again. When we've done different cost modellings, I can show you if you ever wanted to come to the stand and have a look, we're really looking at trying to reduce footprint that you need in cleaning space and reduce the need for scale out, right?
10:19
So the activity is done in one hour, you move on to the next.
10:23
But in terms of this piece of equipment, it's essentially an automated magnetic separation system.
10:31
Simply put its pump tubings in a magnet.
10:33
But it's really clever.
10:34
You write the scripts however you need will help process develop with you and off you go.
10:48
This slide is looking at isolating the cell population that you want.
10:51
And we do that with something called Dynabeads.
10:55
Essentially, it's a magnetic bead attached to an antibody that they raise against different CD markers that you may or may not be interested in.
11:04
And it's like the concept when you're at school, right?
11:07
You get standard iron filings, you run over the with a magnet and, and pulls the iron out.
11:11
Now it's the same concept here, right?
11:12
We're trying to pull the cell population out that you would like.
11:15
So that's the cell isolation kits.
11:19
And then with the cell depletion side is the other part, right?
11:22
We don't want to be putting magnets inside of people.
11:24
So when those detach either actively or passively, whole other ball game that I won't get into today.
11:29
But there are different ways in how they'll come apart.
11:31
It's a way of being able to separate out those systems.
11:33
Same piece of equipment.
11:36
And the cell isolation kit is one step, one litre is the whole volume because that's the maximum amount you can get on the magnet at the time.
11:49
But when it comes to cell separation, that's a really continual processing.
11:54
It's not clear on this at all.
11:55
But there is a serpentine bag, right?
11:57
So it forces your product to go through different channels.
12:01
It pushes it over the magnet as many times as possible.
12:03
And that's automated with the pump.
12:05
So by doing that, you've maximised the surface area you collect, but you can also keep processing for whatever volume you have.
12:14
So we've tested as little as 10 mil.
12:17
The highest we've been so far has been 10 litres of cells.
12:21
And we've used a whole 8 vials of Dynabeads, which is ample.
12:26
We may increase further in the future.
12:28
It depends if people ever end up going to that volume.
12:30
I mean, it may do eventually, but at the moment that's more than enough and we'll see some data over the next couple of slides.
12:37
I think it's the next slide actually around residual beads.
12:41
But in total we tested up to about 50 billion cells and the system has worked really well all with good residuals, which I've just very quickly showed here, right.
12:51
So the DynaMag is the previous iteration.
12:53
So DynaCellect is the piece of equipment I've just been showing.
12:56
That's a nice automated closed system.
12:58
DynaMag is also a closed system, but it's very manual.
13:01
You move about and that there is a bit of operator variability, which is why the there's such variability in the left-hand side.
13:09
When we go to automate it, we're moving that operator variability and by creating these different channels, we've been able to get more robustness, more consistency and way further than what the general accepted limit of about 100 beads per 3 million cells is.
13:24
So I'm going to be quiet in a moment and let Suzy actually talk about the fun QC stuff.
13:31
I just wanted to highlight a moment that there is also all the complementary analytical side of parts that Thermo Fisher can help develop with, whether that's the, the equipment or the actual assays itself.
13:44
And this final part for me is just trying to break down the different steps and show the different products that are very quickly highlighted.
13:54
I'm sorry it's been such a quick tour around, but kind of explain where they fit in for the processes and just to highlight that QC is going to fit all across the board and also as a cue for me to be quiet.
14:06
So I will let Suzy come take over now.
14:10
Happy to answer any questions afterwards as well.
14:12
But if any of you want to look into any more detail, we'll be about on booth 10.
14:21
Great.
14:22
Thanks, George.
14:22
Oh, that's quite loud.
14:25
So yeah, on my turn now to take you through how to further reduce your contamination risk by implementing some of our rapid solutions that we have for mycoplasma and sterility testing.
14:37
So we understand that cell-based therapies are a very challenging product for you to manufacture.
14:42
They're likely therefore to have some very key considerations when you're deciding which analytical assays to implement in order to aid you in your manufacturing.
14:51
So you need to look for solutions that will either meet or exceed those regulatory expectations.
14:57
You want solutions that are going to be a complete workflow, so taking you from your sample to and out so quickly that you can implement those into your lab for very routine use.
15:08
And finally, any applications that can be scaled, taking you from one sample to many if needed.
15:16
So all cell therapies are going to require extensive characterisation and testing to support their commercialisation as shown on the left side of the screen.
15:25
Some key cell therapy testing that you're going to look into is identity testing which covers potency, viability, titre and purity testing, which includes contaminant testing such as sterility testing, mycoplasma and impurity testing for process related residuals.
15:42
So my talk today I'm just going to focus on two of our products that's mycoplasma detection solution MycoSEQ and our very recently released rapid sterility testing kit SteriSEQ.
15:56
So quickly taking the CAR T therapy workflow as an example, your typical testing points for contamination for sterility testing for the presence of microorganisms will be after final processing at the end there and testing for mycoplasma contamination is typically going to be at the end of your cell expansion.
16:15
If you can introduce rapid nucleic acid-based technologies that will provide same daytime to answer, then you can now establish additional testing points across that whole workflow as a contamination risk mitigation strategy.
16:29
So you can test the raw materials that are coming in at additional in process points as well as those required kind of lot release stages.
16:37
This will mean obviously you can detect a potential contamination event sooner as well as possibly help identify the root cause or where any contamination has come from.
16:48
And obviously ideally all those in process tests are going to come back negative and that's given you complete confidence in the product that you're making before it continues forward.
17:00
So I think mycoplasma testing is quite a good example of where we've seen a move from more traditional cell-based assays to rapid PCR based methods that have been successfully adopted in the industry and that's been in line with regulatory guidance.
17:15
So the European Pharmacopoeia Chapter 2 point 6.7 since 2007 included the option that you could introduce a nucleic acid application technique as an alternative to the 28-day cell culture method following suitable validation.
17:31
So this is really meant that today rapid real time PCR methods are globally accepted for the testing of Mycoplasma at lot release.
17:39
And that means that we've been able to move away from those very long testing samples, which could delay the release of material or even the need to work with live mycoplasma and specialist labs or contractors and instead move to more in house rapid qPCR assays with that same day time to results.
17:58
So our MycoSEQ solution, which fulfils this qPCR based method includes a full sample to workflow solution.
18:06
They're off the shelf kits that are designed to meet those regulatory expectations around validation.
18:12
So sensitivity and limit of detection.
18:15
It's also supported by integrated software with 21 CFR part 11 compliance instrumentation, computer system validation services as well.
18:27
This slide actually shows the system.
18:30
It's a fully integrated real time PCR solution.
18:33
The latest kits that we have is called the MycoSEQ Plus assay.
18:37
This is a highly sensitive real time PCR assay that uses TaqMan chemistry to detect down to 10 colony forming units or 10 genome copy equivalent per mil as expected by the LOD from the European Pharmacopoeia.
18:52
And obviously we've tested that on those species detailed in the Pharmacopoeia as well those panel of Mycoplasma species themselves.
18:59
We have we can detect over 200 different Mycoplasma species and no cross reactivity to closely rated bacteria.
19:08
The assay itself being a PCR based workflow takes about 5 hours to start from your sample, extract your total nucleic acids, run the real time PCR assay itself and then assess your data.
19:21
So this allows you obviously to detect any potential contamination very quickly and even expedite the release of products as well as implementing in process testing for risk mitigation strategy.
19:33
A really key feature to this kit is our unique discriminatory positive control.
19:39
So this is used to confirm extraction and PCR performance without the risk of cross contamination.
19:45
So it has its own independent signal that is different from a live Mycoplasma true contamination.
19:52
We also have an internal positive control, again a separate signal that proves that your PCR reaction has worked.
19:57
So no false negatives.
20:00
Data analysis from the workflow is all supported by our AccuSEQ software.
20:04
So this has security audit and E signature features and also automated presence absence sample calling for very simple data analysis.
20:17
This slide is a quick overview of the nucleic acid extraction kit, the PrepSEQ chemistry that I just mentioned.
20:24
So this is one of our kind of more universal sample preparation kits that is used not just upstream of our MycoSEQ for mycoplasma detection, but actually for our residual host cell DNA testing range and residual plasmids as well as some of our viral titre kits.
20:39
So it uses magnetic beads to extract 2 total nucleic acid.
20:43
So it'll work for double stranded, single stranded DNA and RNA, obviously single stranded RNA and is suitable for those mentioned applications as well as working with high recovery and very complex sample matrices.
20:58
The slide here is basically showing you that it's quite scalable in the sense that you can take this kit and use it manually for a very low throughput, say up to 16 extractions.
21:08
Or you can use it very highly automated on our AutoMate Express in the middle, which is a completely hands off system, taking you from between 1 and 13 samples per run.
21:17
Or you can go through to very high throughput on the end, which is our KingFisher Apex, which is 96 extractions in one go.
21:23
So thisis just saying we've got the scalability covered in this aspect.
21:29
OK, moving to sterility testing, so obviously it's a similar challenge in sterility compared to the mycoplasma.
21:38
So you're required to perform sterility testing.
21:42
But again, the challenges come around having a short shelf-life products, very small production volumes and typically the compendial method is a 14 day cell culture and either other rapid growth based methods still take a minimum of five days.
21:58
We have seen in the literature now that from the regulators around an alternative risk-based approach.
22:05
So there's guidance from regulators that you can use in process testing as part of your strategy for checking no gross contamination event.
22:19
It's not necessarily a lot release method, you still need to put form the final lot release, but you can follow and be confident up to that point that you have a clean product.
22:29
So we're seeing the regulators moving towards that strategy which is really positive.
22:35
And obviously, again, a rapid qPCR based method will fit here as a really nice solution to enable you to do that in process testing.
22:43
Similar concepts as well in terms of the compliance side of things, we know who we're speaking to in terms of QC and GMP labs.
22:51
So I'll just show you our SteriSEQ solution.
22:55
The workflow after you've extracted your DNA during sample prep.
23:01
Again, it's an off the shelf kit for sterility testing.
23:04
We're using the same Taqman chemistry.
23:06
This is a highly multiplexed assay.
23:09
So what's different about this kit is that we are simultaneously detecting bacteria and fungal samples in the same well.
23:17
And that's why we need that multiplexing.
23:19
So we have different signals for different targets running on our QuantStudio 5 real time PCR system and that same AccuSEQ software.
23:29
So all the same features as well as inbuilt templates to allow you for automated presence absence calls.
23:38
The design of the kit is against the 16 SRRNA for bacteria and the 18 SRRNA for fungal.
23:47
We can detect down to between 5 and 25 genome copies of the assay, very similar 5 hour process.
23:55
The discriminatory positive control aspect has been designed into this kit as well.
24:00
So you have something that you can spike into your sample matrix to check that you're able to successfully extract and that you haven't got any PCR inhibition in that sense.
24:08
And I think that covers everything.
24:11
And obviously the fact that it's simultaneous identification of any bacterial or fungal means that you can use less of your sample product and you know, hopefully save on cost as well by not repeating assays.
24:23
I think that's covered all the main points.
24:25
So I've obviously introduced just two solutions today that will support cell therapy manufacture, but we do have a host of other applicable analytic solutions which are shown here.
24:35
So I'm just going to quickly call out our microbial identification system for environmental monitoring on the end there.
24:42
So that's a product that we have for the identification of bacterial and fungal species.
24:46
So typically more a kind of a clean room support system there for environmental monitoring and using sanger sequencing and electrophoresis.
24:54
And, and the other testing solutions are particularly for those working with viral vectors is more around, yeah, residual DNA and viral vector characterization.
25:04
The lady is standing up.
25:05
So I think I'm pretty much done.
25:07
I'll just global support.
25:10
We're a very big company.
25:11
That means that we do have a lot of people able to support you in this process.
25:14
So obviously field application scientists like George and I are there for more Technical Support and training.
25:19
We do have obviously our engineers, IT support for computer system validation and regulatory expertise as well to really help you with that journey.
25:28
And I'll finish.
