"Hello and welcome to Sartoria's webinar today entitled 'Robust by Design Re imagining Bag Technologies for Bulk Drug Substance Management' presented by Marion Monstier and Stefan Woodhouse. Please allow me to introduce myself. My name is Rachel Wilkes. Hello and I will be your moderator today.

Let me now introduce our speakers.

Marion Monstier is Global Product Manager for 3D and 4 Technologies.

Marion earned her Master's degree at the Biotechnology Engineering Institute of Bordeaux in France and is now the Product Manager for Freeze and Thor Technologies.

She joined Sartorius in twenty sixteen as a Field Application Specialist for the fluid management technologies before transitioning to the product management role, First focusing on fillfinish technologies and then transitioning on to freeze and thaw technologies.

Stefan Woodhouse is our product specialist for freeze and thaw technologies.

Stefan earned his PhD in Biochemical Engineering at the UCL in London. He joined Sartorius in twenty sixteen as an Application Specialist for Fluid Management Technologies and now provides support for the implementation of the CELLIUS freeze and thaw technologies throughout Europe.

So, let's get started. We hope you will enjoy today's webinar about freeze and thaw technologies.

Marion, Stefan, please begin when you're ready.

Thanks Rachel for the introduction.

Now, the reason why we're all here, freezing and thawing of bulk drug substance is an incredibly common thing within our industry and we have discovered solutions for doing this.

And this ranges from individual single use bags, which of course appeal because they are ready to use, but they are of course brittle and they can develop micro cracks and you can have leaks from that. You can also have bottles, which are very common as well, but these can have problems with their cap closures and maintaining sterility. And then finally on the single use side, you also have bags that can be assembled into protective frames. But then you also have the problem of that burden being on the operator to assemble this and then there are also risks of leaks and issues.

And when we're talking about this sort of process, so freezing of drug substance, even a small leak can be unfathomable amounts of money, as I'm sure everyone's aware.

Now, this is all well and good, but as we become increasingly globalized and manufacturing networks expand, we end up with more and more pressure being put on freeze and thaw operations.

Now here we start to encompass multiple steps. So it's not just the freezing and thawing of bulk drug substance, but it's also the shipping and transportation of it and how to fill and all these other points. And what we end up seeing is that we end up with a problem with existing technologies. So legacy stainless steel or existing single use technologies really have limitations when it comes to scalability, logistics and safety.

Now, what we want to do is build a RS which addresses all of the issues that we see and describes all of the important requirements from the industry and most importantly from customers.

And our focus really is to build a strong collaboration with end users that meets all of these requirements.

But how do we go from a URS to a solution?

Marielle? Yes, certainly. That's a good question, Stephen.

Indeed, this exercise has required several expertise. I would say first the material expertise and the knowledge of characterisation methods for the different materials to be chosen. This has been really a prerequisite for the design and concept phases.

Then some design science is required as well in order to understand the impact of different designs versus a specific application and ultimately adapt the packaging elements accordingly. And of course, the third one is the knowledge of the environment, meaning the expertise around the presence of processes, presence of applications.

And to summarize, these are really the important pillars to build a reliable all in one solution.

So sorry, yep, looking more into the details.

The first thing I mentioned is material expertise. This corresponds to really the knowledge and characterisation knowledge of the interesting properties of materials.

What we did is to characterise candidate materials at different low temperatures in order to inform the design in early stages, meaning really understanding micro and macro properties changes with the temperature change.

The second thing is really to understand the environment, because this will drive the final design quiz as you will likely adapt the design according to the application. So this corresponds basically to the understanding of shocks, vibrations, temperature and any external parameter that has an impact on the potential stresses that can experience the product.

And finally, the design expertise leads to the design definition and its final packaging. And that is possible with the use of simulation tools in order to simulate different designs according to the application and also in order to adapt the design especially to reduce stresses to identify most sensitive areas.

Concretely, to build a reliable tailor made solution as we mentioned?

We will go through our first call, because we wanted to ask you the question. How would you you imagine you have to build a product tomorrow for your 3DENCEO applications, How would you exactly do it?

There are several approaches to develop a product.

Would you take standard materials from a database that you have existing internally, materials that you know.

You would build multiple different prototypes from this material database, test them in parallel and choose the best one.

Second approach would be like you take those standard materials, you do a first prototype that seems to be the best one, you test it, you modify it, you test it again, and you do several iterations until you reach the perfect design.

And the third approach would be, as wind hoarders a little bit, like you characterize material candidates as if you know nothing about those materials.

You simulate designs.

You do a fast prototype that you test based on these simulations and all of the materials you have characterized previously.

I don't know if the poll is working. I'm asking the moderator now. Okay, seems to work.

So basically those are those three approaches and to be honest there is no perfect answer. It's just really think how would you do it if you were to do it yourself.

You don't have any supplier, you need to do it yourself.

Okay, I will give you a few seconds to complete.

Again, there is no perfect answer or yeah perfect approach.

Okay, so yeah seems that most of you would do several iterations or do also multiple prototypes in parallel. And less of you would do the full characterization, simulation, and so on. That's very interesting actually.

Okay, thank you for answering.

What I will describe is actually most likely the third option, which is really what we have taken as approach at Sartorius. And let me explain you why.

I would say if I need to explain how to move from a concept to an actual solution, and a solution that can be used for GMP manufacturing, I guess one way to explain is to use a typical quality risk management approach, where usually you would identify and rank risks and then define preventive and or corrective actions that are then controlled with a control plan.

Right? So basically the process looks very similar even when you develop a product.

The first step is to have this concept based on the important pillars I have presented in the introduction.

Because that's a little bit your risk analysis, you identify where you have risk in the application and you design your product accordingly.

And this concept or design addresses really all major risks identified linked to this application.

And this final concept will be tested, second point, extensively in initial validation. And third point, once released and produced in routine, process controls are performed to ensure product profitability and therefore robustness in routine.

So let's deep dive onto the first one a little bit more in two details.

As I introduced previously, material characterization is very important, really to select the materials that demonstrate best performances against the application.

In our case, of course, the application is related to low temperatures first. And when considering single use systems, it is important to consider not only the film or the tubing, but also protective packaging elements as well as engagements, connections and so on.

You can see here some examples of tests that our engineers are used to perform to test different parameters of the materials.

The list is not exhaustive, but I have chosen some examples.

So basically, tensile strength corresponds to the resistance of a material to fracture when stressed.

The impact resistance is more characterizing the extensibility properties, so basically what we could call the thin strengths.

The burst resistance, which I think is probably the easiest to understand, and cold crack corresponds to really the characterization of the material transition between ductile and brittle behaviors at macroscopic levels.

And all of those cells are really here to help identifying the most adapted materials for the different parts of the product to be designed.

Once those materials are chosen, then moving to the design and conceptual phase. Prediction and simulation tools are really useful in order to evaluate and understand the container behavior into representative testing. These are some examples of static and dynamic simulations, which allows to avoid multiple design iterations, with really dynamic simulation of stresses and shocks experienced by containers during handling, shipping. So you see here, for example, the effect of filling, the effect of freezing, or the effect of different shocks.

And at the end, the result is that we get a final design after the simulation on which we have a high confidence into the robustness and performances against the application.

So this design is really what we call tailor made then for the specific application, in our case freezing and sewing, including frozen shipment as well.

The result for us, so this is the schematic representation of our resulting concept at Arctorius.

So our concept is really a bagging plate concept, which we can call a kind of cassette, which includes a flexible bag. So the bag which is really at the middle, which will contain the track substance, made with one of our film.

And this bag is kind of sandwiched between two semi rigid polyester plates and a surrounding protective rigid shell.

This container, once fully assembled, is really delivered preassembled, sterile, and ready to fill.

The plates, so the sandwich plates, secure the bag and lines, but the role of these plates are really to add impact resistance, suppress wrinkles of the bag, restrict the degree of freedom of the bag so that the bag does not move during handling and shipping that much, and it also reduces ice bulging formation due to the ice that is shrinking when it's forming during the freezing process.

It has been really designed in order to protect the film and bag from the exact stresses that can happen during a freelancer process.

And the surrounding shell is designed to bring maximum robustness to the assembly, protecting from potential shocks and also to stack containers on top of each other.

But Marill, how do we actually prove robustness? Because we've talked about it from a validation perspective and all of our initial upfront design and testing. But how would we actually go about proving this at all? And this is also for the audience as well.

Would we reuse existing tests that have proved reliable in the past? So these are the existing traditional drop protocols, just impact tests, that sort of thing. Will we start from the ground up? Will we develop an entirely new basis for testing or would we want to really prove how strong our bag is by then pushing it to its limits?

So really dropping it from a height, jumping on it, etcetera. Some really good demonstrations of strength.

So yeah, as I was saying, we've got the three options here.

They're all valid in their own right as well, but realistically here we're looking at very traditional methods which have been used for a long time.

We have a completely ground up approach. So starting from first principles and building a new testing method, or really pushing the bag to its limits, really getting in there and making sure that no matter what you do, it will not break.

I'll give everyone a chance.

I see. Okay.

Interesting. Mario?

Yeah.

It's interesting because it seems that we have a very large audience who thinks differently actually, but most of you, a majority, is still thinking that okay, let's use what is already there, conventional drug testing basically.

That's very interesting. I was actually thinking that most of you would push the back to its limit or things like that.

And I want you to give some examples of if you would do that, what would it give if you think about phones, for example? So if you imagine doing thing with a phone, it will probably look like this.

So you would do, like, crazy tests to test the limits of the of the phone. Will it resist to cutting with knife or things like that?

Basically what we want to explain here is that the assurance of robustness is not about dropping a product from ten meters and show that it is not broken.

Sure it shows that it would be robust, but there is no rationale behind this test and there is no rationale related to the life cycle, the real life cycle of such product.

So it's like cutting an iPhone with a knife. Who would do that in the real life? No one. So that's not realistic and that's not connected to the real life.

So proving robustness really requires deep application knowledge, meaning really the understanding of the product life cycle, how is it used, in which environment and so on. And based on this experience and this knowledge, then a protocol for testing can be built defining worst case scenarios of those real life cycle use cases.

So that is also what we did. It means that if we go back to our logic flow, once you have your most adapted design with these simulations and so on, how to test it in a way that is representative from real life? This is with basically product and process qualification of course.

So what we did is actually to rebuild entirely the basis of our qualification protocol. Why? Because there are lots of standards existing on the market and St. Troyes knows also how to test single use bags, of course. But there is no true existing standard describing how to test freezing containers in terms of robustness.

And there is a lack of data to quantify shocks during manual handling of such frozen containers.

So Sartorius engineers worked on a preliminary ergonomic study together with an external packaging institute to quantify those strokes and build this database. And this database was then used as a basis to quantify, to define the protocol, meaning what tests, what drops to perform in order to reproduce the same shocks and peaks, let's say, that were identified in the preliminary ergonomic study.

And these protocols, these methods were then introduced into a larger protocol covering all of the other steps of the life cycle of freezing containers, meaning including filling, shipment, draining and so on.

Of course, I cannot share the entire approach into details today. This will take too long, but in case you would like to learn more into details, this is something we can discuss offline as well.

But basically this is the entire life cycle I was mentioning, where you can see all the steps freezing containers can possibly go through. So it includes handling tests, as I described in the previous slide, shipping tests which are performed according to a standard, the ASTM D4169.

The evolve resonance cycles to also be in worst case conditions for the stresses applied to the polymers, and that mimic also potential partial depletion at the drug product site and refreezing.

It can happen that you are not throwing and draining the entire container and you would use only half of it for example.

And we are not just testing typical case, we are testing worst case for manual handling, case for pallet handling, because we use also the assurance level one of the ISTM start out, which is the most stringent in terms of testing intensity for shipping.

And for each test, worst case conditions are also identified through rationales to determine, for example, the pallet configuration to be tested and so on.

The goal at the end is really to connect those bags to the real world, the real life cycle.

We wanted to give you some examples. So these videos are showing you some examples of the bag drops performed with a specific drop tester that was designed specifically for those purposes.

The goal is to have drops that are performed in a reproducible manner no matter the technician performing it.

So each container was qualified using those tests, which are performed in liquid and frozen states before and after shipment.

The standard protocol basically includes seven drops and those seven drops are performed five times each, which means that in total each container to be qualified and released has been dropped one hundred and forty times.

So at the end of the day, it is not a ten meters impressive drop, but this is a series of drops that are as close as possible from what could happen in worst case real conditions.

I will let the operator end the test.

So you see it's corner drops, sides drops, yeah different basically all sides of the containers and corners are drops.

Okay, and these videos are examples of ASTM tests. You probably know about those tests. So it's just testing the robustness of shippers.

And you need to imagine, inside of these shippers, we have a single use container filled with water, in our case of course, and that we leak test at the end. You can see that the shippers have fallen off.

Can tell you that they have survived to this test, but it's not supposed to happen, of course.

Wow. So what you're saying in the end is that our containers have gone through one hundred and seventy containers have gone through the entire life cycle tests that we've developed in the past, but then also as you'd expect that we've also looked at it from a shipping perspective. So we've done the ISTA characterization for the thermal performance and then the ASTM performance for the mechanical robustness. But okay, we've done all of our validation work. That's fantastic. But coming back to our logic flow, Sorry.

Coming back to our logic flow, we have now gone through the design and we've validated the bags to make sure that they perform as we expected but we need to also make sure that what arrives at the end user site is always what we've done all of our testing on because there's no sense in going through all these design steps and doing all this validation if the product that we've tested isn't what arrives. So in terms of standard process controls, we need to really make sure that we have a sterile product that arrives because there's no sense in having a single use product if it's not sterile and ready to use. But we also need to make sure that it's integral because again, there's no sense in having sterility if the product's not integral in the end.

So talking about integrity of bags, what frequently gets discussed is leak tests. Now all of our bags get submitted to leak tests.

Now these leak tests, they detect leaks above a hundred microns. Now a hundred microns may seem quite small but what we call this is really a gross defect. This is really for us to quickly discard any bags that have had manufacturing errors or had something go wrong in production.

But what we're talking about now is integrity. And what Sartorius has developed is a supplier integrity test.

Now the reason we can call it an integrity test and not just a leak test is because it's actually correlated to microbial ingress.

Because like with all of our validation tests, there's no sense in doing all these tests unless they are actually correlated something meaningful.

And what we have demonstrated is that leaks below a, or holes in a bag below two microns will not actually allow any microbial ingress because it is just too small a defect.

And the helium integrity tests that we have will detect leaks from two microns and above.

And given the criticality of freezing operations and given that we are talking about drug substance here, it's of course important that all of the bags are tested to make sure that every single bag that arrives at the customer site is integral.

But we don't, of course integrity is one part of what we're doing in terms of our process controls and our testing, but there are other points that we need to cover. We also of course make sure that things are sterile. We perform bacterial challenge tests. We do routine batch testing for endotoxins. We have our own particle prevention program. All of these either use representative samples or continuous tests of our products as they come away. And these are of course all summarized in our certificates of release so that every time one of our extensively tested products arrives, that you have a full overview of what testing is being performed and what our standard controls are.

The result of this is that, as we discussed now, we have all the testing gone through to make sure that what you have is being tested from a real life cycle perspective. So we have performed handling qualification. We've made sure that we can go through multiple freeze and thaw cycles and that the freeze and thaw cycles are characterized.

We have done shelf life studies and stability studies.

Really, really importantly, as we move into this more global network where different sites, the manufacturing sites and the filling sites are on separate locations, shipping becomes more important. And we have done that testing as well using the standards of ASTM and ISTA for mechanical and thermal performance of shipping. And we've done that characterization work.

And finally, course, we're not just talking about frozen product in the end. It also needs to be thawed to be used. So we've also done some thawing studies and generated thawing profiles.

So from a use perspective, all of this work will then alleviate your own validation work to make lives easier. But as I've been mentioning before, it's not just the test we've done up till now, it's also the fact that we're making it truly ready to use by ensuring that every bag that arrives is equivalent to the bags that we have tested. So we have those in process controls, that in process monitoring to make sure that everything that actually arrives is what we say it is and that it will meet the standards that we have created and tested.

If that's not enough though, because we are aware that there are customers that have very specific conditions that we can't take into account every possible outcome. So we also offer additional support to customers through our confidence services. Now these services can do real life shipping validation. So if you take the ISTA and ASTM standards, yes they cover the majority of eventualities but sometimes there are longer distance shipping routes. Sometimes there's a risk of a container getting stuck in customs somewhere or there's particularly bumpy road that you know about coming up to your production site that you're concerned about.

These can be then tested and reproduced.

We can also go the other way and look at it purely from a simulator perspective. We can redo some of these tests but with slightly different conditions.

We can also look at chemical compatibility because of course we have gone through all the standards and have done our best characterize the film fully.

But if have a compound, for example, that has a particularly odd or niche compound in order to keep the product stable, we won't have a record of that. But what we can do is that we can provide a study in order to characterize the film in contact with that compound to then make an assessment on your behalf to see the chemical compatibility and that no harm should befell well, our product but more importantly, your product.

Integrity testing will also be an additional service that we can find, so additional integrity testing, I should say.

And finally in terms of extractables and leachables, this goes back in a way to the chemical compatibility side where we're not just talking about the impact on the film and what will happen there but also what will leach or what can be extracted depending on the conditions. But in real life conditions, what will leach from the film into your product when put in real process conditions. So that's something again we can help with.

So with all of these things said, what we're doing is we're offering a solution that will go from zero point five liters working volume up to twelve liters working volume across four different bag sizes in a bag that, as Mario has described, consists of a bag within a cassette within a shell.

This combination allows for a unique set of features because not only is it preassembled and so ready to use and easy to use, but we also through this unique design have great product visibility. There's no need to disassemble the cassette in any way in order to see the product. It can be immediately seen through our SafeCore plates. These plates are also of course easy to wipe down because they're flat.

As we've already discussed from a validation perspective, we've gone through multiple freezing for cycles all down to minus eighty degrees because minus eighty is the standard that we see for drug substance.

And the actual design itself allows for multiple different features to be integrated that wouldn't be possible otherwise. Most crucially, I think, is the tubing management because what we want is to enable the tubing to be easily accessed because that is of course how you're connecting to your other process steps.

But with that easy access, normally you risk a loss in robustness because the tubing is the weak spot of the assembly. But what we have here due to the design is we have tubing holders integrated around the perimeter of the bag that are easy to access but also hold the tubing in place to protect it. And then a few additionals are the fact that we also have the option of adding a tailgate sample to the bag, that we have a place for labeling and that there's also space for any connector that you really desire.

So when we're talking about take home messages for you, we're saying that material science and application expertise lead to reliable and tailor made solutions for you and that assurance of integrity is guaranteed by having an initial robustness validation process. But most importantly, it's verified by routine process controls.

And that with all of this together, that you can really simplify and really alleviate your validation workload by relying on the expertise of your suppliers. But with all of this said and done, I think really the proof of robustness isn't what we can say and can demonstrate through all our tests. The real proof is that over fifty thousand of these bags have been trusted to deliver sixty five billion doses of vaccine. And I think really that says it all about the robustness.

Thank you for listening everyone. We really appreciate your time and as previously mentioned, we are very happy to take your questions now.

Well, thank you, Marion and Stefan for that very interesting presentation. We have had a couple of questions come through. Please do keep sending them on to us and I will read them out. Here's the first one: How do you ensure flexibility of the design in adapting connections or tube types when proposing a ready to use solution?

I can take that one.

So I would say there are many ways to qualify the word flexibility, but if we look at the possibilities to change tubings and connections or the design, it's true that we have a design scope to keep the tubing in this integrated tubing order, for example, keep the connector under the protection of the shell.

But we do have possibilities to customize, to change the connector type, to change the tubing type, as long as it stays under the protective umbrella of these plates and shells.

And that is possible with we have done several design checks, for example if you have a bigger connector type that you are used to use for your process, this is something that we have checked so that it fits into the shell. So most of the standard collectors used and types of tubing used on the market are possible as long as they are validated for a free dental application I would say, meaning that they won't break at minus sixty, minus seventy, minus eighty degrees of course. And there are also ways to customize, for example, if you want to manifold containers, like five containers, you want to link to each other so that you fill them all at the same time. That is also things that are possible with designs.

Thank you.

I had a couple of questions come in about the testing and I think it might make sense to answer them together. The first one is whether the testing is done in each and every single one of these freeze and thaw cassettes. And the second question is saying that the tests do not seem to be repeatable and is there a possibility that a robot can be used to perform these tests?

Thank you.

I will also take that one. So to answer to the first part of the question.

So we are talking about initial validation. So in initial validation, meaning really when we designed this product to release it on the market, we have tested each and every cassette, meaning, Stefan mentioned, bags, mid sized bags, large bags. All of those before being released have been tested through the same method, same protocol and most likely by the same people as well.

We don't use a robot yet. We have tried however to indeed make this test as reproducible as possible.

I've shown you in one of the videos the drop tester that has been designed specifically for those tests. On the drop tester what you cannot see on the video, have defined length, defined dimensions, heights for how to perform each drop. So it is still performed by a human, by an operator, but I can tell you it is very reproducible thanks to this drug tester because there is no way to drop from a different height or different distance or anything, because this is all defined by this drop tester.

Maybe in the future we can think about robots to test those containers, that would be great actually. But we also wanted involve humans to introduce human errors potentially as well.

Not in the way things are tested, but basically when you use those bags, this is an operator, a human transferring the bag from the freezer to a shelf to the next step. And we wanted this to be done by humans as well. So yeah, I would say this is probably even better to have humans testing those bags.

Thank you. Thank you very much. Here's the next one. Is chemical compatibility being done in liquid or frozen state?

Would you like me to or would you like to?

As you want, chemical compatibility.

So actually it depends what we're talking about, whether we're talking about the initial studies or latest studies, because I think Marielle can comment on the initial studies. But what I was talking about regarding confidence studies, the confidence studies that we offer can be done either way. The idea is that we offer you the option of whatever your process conditions are, that we reproduce them in order to discover the compatibility. So this will be anything within the operating parameters of what the bag is designed to handle. And those will be with various concentrations of each chemical or compound and at various temperatures. So it depends entirely on what the customer requests.

From the initial testing, well, I believe it was in liquid format, but yes, I need your confirmation.

Yeah, I can comment if you want. Usually initial validation for chemical compatibility or even extractables for example, we do that in liquid first, but also at higher temperatures because higher temperatures will always, let's say, put the multilayers in worst case conditions. So when looking at extractables, for example, you will always get more extractables at forty degrees compared to zero degrees.

At lower temperatures everything is slowed down basically, so we usually perform t"