Hi everyone and welcome. My name is Lizzie Tian and I will serve as your moderator today. Thank you for joining us for today's webinar development of innovative single use file processing film for strength and leak resistance. As your moderator, it is my road to ensure that we make the most of your time with us. I'm here today with Shannon Cleveland. Shannon is a Research and Development manager and technology lead of Ultima's Film. She started at our company in 2006 and has a long track record of technology accomplishment. Having contributed to multiple successful filtration product development since 2018, Shannon has been essential innovator within the single use and integrated systems R&D team, contributing to breakthrough film and single use bag improvements. She holds a bachelor's degree in Chemical Engineering from Northeastern University in Boston, USA. Before I turn things over to our presenter, I'd like to cover a few housekeeping items. At the bottom of your screen are multiple application widgets you can use. There you can also find our reaction button, indicated by the thumb up emoji and allows you to give immediate feedback on the present presentation topics or anything that stands out. All the widgets are resizable and movable, so feel free to move them around to get most of your space. You can expand your slide area or maximize it to full screen by clicking on the arrows in the top right corner, but if you have any questions during the webinar, you can submit them through the Q&A widget. I'll try to answer all these during the webinar, but if a more detailed answer is needed or if you run out of time, it will be answered later via e-mail, please know we do capture all questions. You will also find the opportunity to participate in a couple quick poll questions throughout the session. I encourage you to take part in these surveys. If you're watching this webinar on demand, you can still submit poll questions and responses via the Q&A widget. The webinar is being streamed through your computer, so there is there is no doubt in number for the best audio quality. Please make sure your computer speaker or headset are turned on and the volume is up so you can hear the presenters. An on demand version of the webinar will be available after and can be accessed using the same link that was sent to you earlier. Lastly, attendees who wish to receive a webinar certification will need to fulfill the criteria of 30 minutes viewing time and completing 2 poll questions within duration of this webinar. So that is from my end. Shannon, turning it over to you, all right. Thank you, Lizzie. Hello, Thank you for attending today. First, I would like to cover some background on where the bioprocessing film is used, why the new film is needed and some basics around how we measured success. Single use films can be used throughout the unit operations of bioprocessing. Here's an overview of a MAB workflow. Single use films are used in our Mobius bioreactors, flex ready flow pads, mixers, storage systems and transportation bins. Top industry concerns for single use films are leaks and cell growth compatibility. So before we head over to our next session, let's take our first poll question here. Where are your most concerned about leaks in bioreactors mixing transportation and storage in upstream processes or transportation and storage in downstream processes? Let's keep our audience some time to answer. Thank you for submitting your answer. Now I'm handing it over to Shannon. When talking to our customers, film durability can be vital to their process. The average of just one leak can range anywhere from 50,000 to $600,000, with the costliest leak occurring with transportation of bulk drug substances. With this in mind, we needed to reimagine the film to create a more durable structure. Another area of importance is compatibility with cell growth. Customers can observe variable cell growth that can change by a film supplier and even between lots of the same film. A study published in the Journal of Pharmaceutical Science and Technology discovered that the antioxidant Ergophos 168 degrades during gamma sterilization into a chemical called BDTPP or BIS 24 Dieter Butyl Phenyl phosphate. This chemical was identified as an extractable material that can negatively impact cell growth. We needed to take care when selecting the right resin to minimize extractable leachable profile and to maximize cell growth. It may sound simple, but in addition to developing a durable cell growth compatible film, we also needed to consider other performance criteria that are essential to meet. Some of these criteria are listed here. Bioprocessing films are multi layered to meet the multi faceted performance criteria. Each layer carries a purpose. So where do we start? We started by reviewing the performance of a typical single use film and breaking the structural layers down by purpose. The design for the new film needed its layers to robustly ensure quality performance for all design criteria. We had many tools to help us explore and measure success. To evaluate designs for technical feasibility we used an in house pilot scale five layer extruder laminator that allowed us to screen resins and potential film designs. This in house capability opened the door to engineering innovation, intergroup collaboration and allowed for the creative processes and designed experimentation to be nimble and efficient. The films created in our lab were then instantly available to run a gambit of testing. The resulting performance was then evaluated against commercial single use films. Much of the performance testing was done in our own labs. Shown here are some film based tests such as abrasion resistance with a taper, abrasion tester, flexibility with a Galbo flex tester, tensile strength, tear resistance and puncture resistance using an Instron and one of the film and product tests done externally was impact and vibration testing to simulate transportation. Trial film structures were also tested for TRO cell growth compatibility and benchmarked against cell growth and sterilized glass. The tests were run on our internal lab capabilities. The process starts by making ported film pouches, packaging them, sending them out for gamma sterilization. Cells were then incubated, prepped for culture, cultured for five days, and then cell density and viability were evaluated. First, let's review the development of the fluid contact layer and share the results for its compatibility with cell growth. Our main objectives for the contact layer were to identify a resin without Urca Phos 168 which delivers high cell growth compatibility to use. A resin that could be Co extruded and therefore processed into film. Have a low extractable leechable profile to minimize contaminants in the customer product and provide robust seam strength for manufacture ability when sealing film panels together to create bin bags, mixers and bioreactors. We chose to use a polyethylene resin for our contact layer due to its availability, its stability, chemical compatibility and it's wide range of density and melt flow for processing. Polyethylene also has a history of providing robust seam strength within the polyethylene family. We explored new resin grades, blends and thicknesses of film made from those resins. These polyethylene candidates were evaluated during the extrusion process for its ability to create good film, the durability of that film, the strength of the seal when bonding film to itself, its extractable leechable profile in water and ethanol, and lastly but not least, its compatibility with cell growth. When we identified the best polyethylene candidates that demonstrated processability, durability, manufacturability and cleanliness, we moved forward with testing cell growth compatibility. We created small bags that exaggerated the film to fluid ratio by 7 times higher in comparison to A10 litre media bag or 20 times higher than a 200 litre bioreactor. By exceeding the film to fluid ratio, our studies would ensure that our extractable and leachable profiles would provide the proper environment for the cells to grow at this high film to fluid ratio. We compared cell growth for polyethylene films with and without Ergophos 168 against a glass standard over 5 days of incubation. The polyethylene resin without Ergophos 168 produced high cell density as seen in the green data, while cell growth was poor for the polyethylene containing the the antioxidant as seen in the blue data. Even more exciting was that polyethylene without Ergophos 168 closely mimicked the benchmark of cell growth in glass, which is shown in the graph in purple. This discovery successfully concluded our search for the ideal fluid contact for the new film. The next part of the film development we'll review is the layer that functions as the gas barrier. The gas barrier is an important layer of the film for applications we're maintaining. pH of the fluid inside is vital. Typically for storage applications, ethylene vinyl alcohols are commonly used as gas barrier layers and films. However, they're not known for their flexibility with standard grades. Low oxygen transmission resins, IE good gas barriers, have low flexibility, while more flexible grades of EVOH provide less of a gas barrier. But specialty grades break this trend by providing both a low gas permeability and increased flexibility. To test the flexibility of gas barrier candidates, we melted standard and specialty ethylene vinyl alcohol pellets and extruded them through a dye to form films. In this case, the thickness of these films were five mil. He loaded samples into the Gilbo Flex tester to simulate extreme handling. The Gilbo Flex test repeatedly twists the film over 420° and then crushes the sample. The resulting flex film is then flattened and secured in A-frame with a paper placed underneath. Dye is speeded over the surface. Holes and cracks are detected by dye that penetrates through the flex film. As you can see in the image on the bottom right hand corner, the circled areas of the film are showing where dye passed through the cracks or pin holes and onto the paper. We evaluated the EVOH gas barrier samples after flexing the film 200 cycles. A standard EVOH grade resulted in multiple large holes after Guildville, while a specialty grade robust robustly remained integral in all 100 samples tested. This new specialty resin was chosen for our film because it's significantly improved flexibility when compared to standard EVOH grades, while still providing an excellent gas barrier. In the next few slides, we'll review how we developed a strength layer for the new film structure. This the strength layer was designed to reinforce the film to improve durability. Our team conceptualized how to reinforce the film and drew inspiration from other applications. Much like skeletal structures in animals or veins and insect wings, some man made products like those depicted here have been created with an incorporated reinforcement material to act as a support and boost boost toughness. We enhanced single use film by incorporating A reinforcement layer which significantly increases durability and strength in bioprocessing films and we were awarded a patent to protect the novel design. Finding the right material to provide that reinforcement had the team searching through many possibilities. There were many varieties of material to explore. We refer to those materials as mesh. The variables included. What type of resin would the material be made out of? Would it be polyethylene, Nylon, Polyester, Cyclic olefin copolymer? What format would be best? The spun bond woven, Extruded mesh knits? Would the material have a high density of fibers or would low be better? What would the diameter of the fibers be? Are mono or multifilament fibers the best? There were so many possibilities to explore. It was also the question of where the mesh should function best within the film structure. Should it be on the outside? Should it be under an? Should it be under an outer layer but above the gas barrier layer? Should it be closer to the contact layer and below the gas barrier layer? It was necessary to combine mesh candidates within a film structure to evaluate durability and strength performance. To properly incorporate the strength layer into the film, resins and thicknesses of those resins needed to be added into the experimental design performance of the film. Mesh composite candidates were evaluated for durability. Here you can see some of the abrasion resistance, puncture force and tear strength results. The graph shows nylon in blue and other candidates in yellow. Nylon materials clearly provided the most durability and strength, with the best overall performance and most suitable reinforcement being a monofilament Woven nylon structure. Incorporation of the Woven Nylon mesh with tie layer resins required careful selection and process development along with some new test methods to incorporate the Woven Nylon mesh. There was some additional performance metrics. 1st is interlayer film integrity. This is needed to ensure that all the layers of the film stay together. It needs to remain integral while it's manufactured into product as well as when it's developed. When it's deployed at the customer without layer separation, resin selection considerations included compatibility from the nylon to the rest of the film structure and the ability of the resin to flow in and around the woven mesh. 2nd consideration is film transparency or haziness. An increase to the opacity of the film meant that it would be more difficult for our customers to see inside the bag. For this melt. Flow of the tie layer is important not to trap air as trapped air increases haze and finally the film needs to remain flexible. Resin selections needed to be elastic to move with the film mesh composite. These graphics show how film samples were created on our extrusion lamination equipment and to better describe how the tie layer resins incorporate the woven nylon mesh into the reinforced film. Starting with the graphic on the bottom left, a blown or cast film that was already made is thread below the extrusion dye where tie resin is extruded, sandwiching the film to the nylon mesh. This forms an intermediate film nylon composite. As you can see in the first cross-sectional SEM, the nylon mesh is embedded in a tie resin. On a second pass through our extrusion lamination line, the film nylon composite is merged with another film using another tie resin. The result is a completed film. As you can see on the bottom right, the ample assessment was broken down into different phases. Each phase used test methods to help further refine the film structure. We started with the Gilbo flex test with the same method as described earlier for the gas barrier layer flexibility performance was measured by detecting cracking or pin holes that form during flex testing. During this phase, films were flexed up to 2700 cycles. The eye test results guided the structure development and narrowed resin selections and film thicknesses to eliminate cracks and pinhole formation. After cracking in, pinholes were eliminated, the next assessment phase visually assessed film stress after Gilboflex test. The visual test assessed the interlayer adhesion of the film layers that were exposed to tensile and compressive forces from flexing. This resulted in eliminating stiff tie layer resin candidates, optimizing layer thicknesses, and the reduction of the lamination. Lastly, a haze analysis was implemented. This analysis examined interlayer adhesion by measuring the resulting haze after stretching the thumb. This analysis LED even more film improvements where the formulation was optimized, the production process was finalized, and the interlayer adhesion was optimized. Since we've already reviewed the Gelbo Flex Guide task method with the gas barrier films, I'll move forward to share details around the second phase of the analysis and show how film was assessed visually after 2700 Gelbo cycles. The first step is to prepare the sample so that the visual information about the adhesion lamination can be better quantified. This was accomplished by enhancing the stress using cross polar films along with backlighting. Cross polar films have very fine black lines that run through a film. When light passes through a vertically polarized film with lines running North and South, only those waves of light that run vertically can pass through. When that light then tries to pass through a horizontally polarized film where the lines run east and West, no light travels through. So here you see an image of my glasses where I have a vertically polarized film above my glasses and a horizontally polarized film below my glasses with a light behind that stack up in the areas where my glasses aren't there. It results in no light and gives the appearance of a black background due to the cross polar films. With an object between the cross polarized films like my glasses, the light traveling through the horizontally polarized film is bent and you can see the object between the films. An interesting phenomenon called birefringence enhances areas of high stress where the materials refractive index was altered. Those areas appear as bright, sometimes multi colored areas. These birefringent areas highlight the stress on the lenses where the frames are attached. This concept is applied to post Bilbo film samples and enhances where the film was stressed during flex testing. We refer to these stress areas as white lines. The method increases contrast between stressed and non stressed areas on the film, making it possible to quantify the degree of white lines through image analysis. The second step of the analysis is to process the image to obtain quantifiable data taking that initial cross polar image. This is the process that was used to generate a numerical representation of the film's flexibility. First the image is converted to black and white, then the contrast is adjusted to remove any glare. A macro is then run to binarize the image and remove the grayscale. Then another macro is run to skeletonize the binary image into a pixelized representation of the lines. And finally a branching analysis is 1 aspects of the output like junctions and complex junctions where the intersections of three or four branches correlate to the degree of white lines formed and ultimately the flexibility of the mesh film composite. This method can be referenced in the image processing. The data generated by this method was used to improve the composites film flexibility. These are cross polar film samples between phase one on the left where pin holes were eliminated and phase two on the right where film flex stresses were minimized. Visually, you can see that the stresses to the film after flexing were greatly reduced. The improvements were due to formulation changes driven by the image analysis data. With elbow flex image analysis, we were able to improve the film's flexibility from phase one to phase two by 4X. While the film performance was improved, further improvements could be made that were beyond the sensitivity of the image analysis. Flex test. While flexing the film with elbow, the film's durability is tested with twisting. Durability of the film can also be challenged by stretching the film perpendicular to its surface. Perpendicular interlayer adhesion is measured by the haze that results. After stretching to stretch the film, a blunted tip was created for the Instron puncture test film was stretched using the modified fixture and rather than measuring the force to puncture the fixture would travel a predefined distance to stretch the film. The resulting dimple was assessed for the degree of haze as well as how far the haze travelled away from the point of contact. I will share the progression of haze through all three phases of the analysis. During phase one pilot phase film showed poor elasticity in this direction and not only was there a high degree of haze, but the film also broke. During phase two, the elasticity was greatly improved, no breakage was noted. However, the sample was still hazy after stretching, indicating improvements to the interlayer adhesion were still possible. And finally, in phase three, we achieved good elasticity and lamination adhesion where the haze compared to phase two was reduced by 22% after structuring. Thank you, Shannon. So now it's time to take our second poll question. What do you value most when choosing a new single use film? So this is a multiple answer question so please select all that applies. Thank you for everyone who has submitted their answers. Now I'm handing it over back to Shannon. Now we'll review the outcome of all this work and show the performance of the new strength enhanced film. Here's a summary of the advancements as a result of developing our new film Ultimus. The gas barrier layer has increased flexibility. The mesh provides enhanced abrasion and puncture resistance. The strength layer tie resins provide flexible, robust lamination adhesion. The fluid contact layer is clean, cell compatible, flexible and durable. Comparing Ultimus to five commercially available single use bioprocessing films for abrasion and puncture, the data shows that Ultimus film provides superior performance with 10 times higher abrasion resistance and two times higher puncture resistance. That trend is also seen with tensile strength and flex crack resistance. Tensile strength is 2.8 times higher and 100% more flex crack resistant than competition at 900 Gelbo cycles. All competitive films tested formed holes. While Ultimus doesn't form holes and we know that it was designed to withstand much more than 900 cycles, we ran a study to directly address the high risk transport of bulk drug drug substance. Based on a customer application, we created and sterilized five 1000 liter Ultimus bags. They were vibration tested in a collapsible bin filled with water. The vibration test ran for eight hours simulating 2400 miles of travel distance. No leaks occurred. All bags passed visual inspection and they all passed a post vibration integrity test. All the hard work of developing Ultimus film was effective. Ultimus Films proven durability has the potential to significantly reduce product loss. Ultimus film is targeted to be used in manufactured I'm sorry. The Ultimus film is targeted to be used to manufacture large volume 3D assemblies. Today you can find our new film in storage containers, single use bins, transportation bins and Mobius iflex fire reactors. We look forward to sharing exciting new products soon like Ultimus Mixers and there's always innovation in process to help create new technology and better products for bioprocessing. Thank you, Shannon. Now it's time for our for our now it's time for our last poll question, where would you like to consider Ultimus Film in your process? This is also a multiple answer questions, so please select all that applies. The bioreactor mixing transparent storage with other applications give you some time to answer. Thank you everyone for submitting your answer again. So that concludes our webinar and thank you Shannon for this great presentation. Now it's time to answer a few questions that come in from our audience. But before we do so, I would like to remind you that it is still not too late to submit your questions to us using the Q&A widget. This also applied to on demand viewers as well. We'll try to get through all of these questions, but if we ran out of time, we'll reach out to you separately. As a reminder, this webinar will be available on our website soon as well. All participants will receive an e-mail notification when it's available for viewing. So now back to you, Shannon, for answering the questions. All right, Shannon. So the first questions we received is, have any studies been run to look at performance under cold temperatures? So yes, we have some limited studies where we have looked at the film under cold conditions. Most of those conditions are at 4° C where we have looked at the film flexibility using Gilbo and looking at 900 cycles on the Gilbo at 4C. The film does not form holes at 4C and we also ran some abrasion studies at 4C which also showed that we had an increase to abrasion resistance at that temperature. Also, we've done some testing for transportation in cold conditions. So for Ista 3 E conditions we used A500 litre transportation bin and it was tested for vibration, drop and collision at 4° CI. Don't hear you. Oh, there we go. I can hear you now, OK. Sorry about that technical issue. So the next question is how are the bags integrity tested? So bags of integrity tested after we create the the bag. So after everything has been bonded the ports in the in the seals what we do is we after inspection of all of the seals we actually inflate the bag to a certain pressure and then we look for a pressure decay over a predetermined time that the process is is validated. It's a validated integrity test that we do on on our bags as part of our lot release. We have another question about the white line. The question is, white lines in other films can be concerning. Are you saying that they are not feedback? OK, so I It's true that that white lines in other films can be a cause for concern. However, with our film we've robustly tested the film to ensure that marks on the film are not a cause for concern. We have put this information together in a customer facing document that shows that we've tested the film over all of these different film marks, including white lines ensuring that the film isn't is integral. No, they're not. Concerned. That's great. I'm I'm sure that our audience would love to hear that too. So the final question we have so far is, is the film thicker than the other single use films? And if it is, how does it impact deployment? So, yes, so versus other films on the market, our film is on the thicker side of that range and we have done deployment studies using both our folding mechanisms. So we we will either fold our bags in something we call easy fold where the top of the bag is pushed to the bottom of the bag so that when you deploy from the bottom, it easily deploys. We tested at format as well as that wonderful format which is slightly different also deploys, but that's where our larger formats above 1500 liters and we've done deployment studies, you know under worst case conditions and we've shown that the bag still deploys. So the only difference is it is a little thicker. So the bags tend to be a little bit heavier than some of our other products. All right. That's all the questions we have received so far. Shannon, thank you so much for a great presentation and thank you so much for our audience for all your questions. If we did not get to your question, please feel free to e-mail our presenter directly and to register future webinars or to access our access our archive, the webinar library, please visit our website. You can also download the presentation. Is there another? Question. I think there is. Yeah, I think there is. Oh yeah, OK, there's one more came in just now. What's the validation done during the real transfer transport or only in the laboratory? So we have we've only done lab stimulation versus actual transport. So during like our our transportation customer application study that was done using an ISTA simulation of transportation and for our transportation testing as well, it's all it's simulated, you're muted I think again there you go. OK. If there's any other question coming in, I'll just do a one last refresh here. I think that's all we have. So again, like I said earlier, if we did not get to your question, if you still have questions, please feel free to e-mail our presenter directly to register for future webinars or to access our archive, the webinar library, please visit our website. You can also download the presentation slides into Take Action field that will pop up on your screen once the webcast has finished. So I would like to thank Shannon again for today's presentation. Thank you so much, Shannon, and thank you for our audience for joining us. And have a great day everybody. Thank you. Thank. You. _1732182906469

 

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