Advancing Just-in-Time Release Strategies for CAR T-Cell Therapies

[00:00:03]hello and welcome to this webinar titled just in time release of car t cell therapies hosted by biopharma asia magazine and presented by irving ford head of car t qc laboratories at celgene this presentation will be followed by smart quality solutions for car t product release and presented by laurie dan farmer microbiology scientific director at bio meru my name is stephen edwards and i'll be your moderator before we begin i would just like to inform our viewers that there will be a live question and answer session after both presentations near the end of the webinar audience members can send their questions in at any time during the webinar via the questions tab located directly below your webinar screen and we'll go through these at the end of the webinar now please allow me to introduce our first presenter dr irving ford is currently the head of car t qc laboratories at celgene prior to joining celgene irving was a significant contributor for activities supporting the commercial approval of novartis's car t product chimera irving has over 28 years of qa qc experience in the pharmaceutical biotechnology and cell and gene therapies industries including but not limited to non-sterile sterile and low bioburden manufacturing production irving has designed implemented and managed paperless laboratories irving has a successful track record working with laboratory facilities validation manufacturing regulatory qa r d and compliance personnel to manage and resolve complex quality and manufacturing issues i will now be handing over to our first presenter welcome irving thank you stephen hi as steven said my name is irving ford and i am currently the head of car tqc here at celgene and summit new jersey and today i'm going to talk about um contamination controls testing and release strategy for car t products and first i have to give a disclaimer that the views and opinions within this presentation are those of the presenter and do not represent the views of celgene or soviet's regulatory positions so before i start i just want to give you a little history about cartoon manufacturing so currently there are two products on the market for um cartoon manufacturing they're the viruses camarilla and kites yes carter and a little about these products is that um this is pretty much the last treatment option for patients they have typically gone through three other um options that have failed so this is pretty much a life of death situation for the patient so when we say that we put the patients first in car t we really have the life of the patients in our hands and by the time we get their blood and process and get it back to them some patients have even expired between that time so as you can see um getting products manufactured in back to the patient is very important for the well-being of the patient so because of that i've explored holistic approaches to trying to get the product back to the patient in a hurry from a microbial standpoint because typically the microbial testing takes the longest at the end of the manufacturing process so i have explored ways of trying to eliminate or reduce those time frames so that we can get the product back to the patient so the patient has a better chance of survival so today in my presentation i'm going to give two options for release of rt products the first often is just in time release and the second option is a two-day release so for those of you who are familiar with parametric release you may be familiar with terminally sterilized products that you don't have to do in microbial testing at the end because it's terminally sterilized so that's kind of the equivalent of the just in time release so if you think about parametric elites now we're going to switch the page and talk about just in time release for cart products and for those who may be a little afraid of taking the plant into going into a distant time release i'm also going to provide you with a different option which is called a two-day release so let's get started so in traditional pharmaceuticals everybody when they come to cartoon manufacturing most people either come from sterile or non-sterile so they either fall like nx1 while they follow the non-sterile guidelines but for cartoon manufacturing it's a unique product in that we cannot assure the stability of the raw material that we're receiving because the raw material is actually the patient's blood itself and because of that most car t products do not have a label claim of sterile and as such we are not um subjected to following nx1 we actually are subjected to the atmp got its document and that's what we follow so a lot of rf companies are just getting started so they automatically revert to nx1 but because your product is not sterile in order to follow meet the definition the traditional definition of sterility we go we follow the atmp guidance documents so in the atm guidance documents of course it talks about risk risk and more risk so because car key manufacturing doesn't fall into the traditional way of manufacturing because everything in cartoon manufacturing as of today is mostly manual aseptic manipulations however we're transitioning into more automation but for right now when you think of the process for cartoon manufacturing you think of human beings in our process in our manufacturing process humans are our machine so everything is pretty much done by a person and in order to manufacture in this regards we have to have a risk based approach so i'm going to highlight some of the risk space directions that are given in nx1 so when annex 1 and 2.13 says the risk-based approach is applicable to all types of atm ps and it applies in an equal equal fashion to all types of settings it is highly dependent on the biological characteristics and the origin of the cells and tissues the vectors and the transgenes and i just want to highlight an important thing it says the leveling characteristics of the express protein for gene therapy products the property of other non-failure components the raw materials matrices and the manufacturing process so again here is giving you an idea what you need to look at from a risk-based standpoint remember for car key manufacturing the raw material for us the main raw materials are the main active ingredient is the patient's blood itself the next quest and the atmp guidance document says consider all the potential risks related to the product or the manufacturing process on the basis of all the information available so what they're saying is that you have to take into consideration everything that you know about your manufacturing process and your risk should be based from the incoming materials all the way to the release of your product and finally i'm just gonna take a subset of this the control mitigation measure should be based on current scientific knowledge and the accumulated experience and it says ultimately this evaluation is linked to the protection of the patient so once again keep in mind that for car t our patients are super super sick they're actually at the point of death this is the last treatment option for them so we want to make sure that we don't have anything in our process that's gonna have the remote chance of introducing any adventitious contamination into the manufacturing process because we're dealing with patients who are extremely sick sometimes the patient has pre-existing microbial contamination in their blood already so our hope is that when we receive their blood we have the necessary controls in place to make sure that that contamination doesn't proliferate and actually kill the patient cells because that creates a greater risk for the patient because the patients are some so sick at times that they can't even receive another apheresis to have their blood taken and because of that we want to have all the controls in place so that we don't introduce any evidence examination such that we can get it back to the patient quick transit in a hurry to save their lives so next i'm going to give a high level overview of a manufacturing process for a car t process so we have our incoming lucas releases so the patient is at the clinic of a hospital they get apheresis then we separate out their t cells their white blood cells and extract the t cells then we have the process of raw materials and consumables then there's always the media involved in the manufacturing process and then you have the actual manufacturing steps and you have the final product so again these are the high level buckets for car t manufacturing and these are the major buckets that we're going to discuss today and evaluating a risk-based approach for just-in-time release and a two-day release of the product from a microbial contamination standpoint so let's start with the leucopharesis product so remember i said that this is the last option for patients the cartoon manufacturing this process they've normally gone through three other treatments and the patient has received other treatments and sometimes the patient is near death and sometimes the patient either has some microbial infection that already exists or when the mucopheresis process happens the nurse of the doctor who are actually doing the process may cause contamination of the patient's blood so from a risk-based standpoint this is one of the most important steps in the manufacturing process of car t because if contamination occurs during this step it's probably going to come into the threshing process and those organisms that may be present from an improper leukophoresis will then get into the manufacturing process and then we go through the growth phase or the incubation phase of expanding the cells which is integral media that microorganisms love that microorganism also has a chance to proliferate and cause the depth of the patient cells so this is a point in the step in the process where you want to make sure that you have some rapid release testing because you want to be able to test the look of reese's material just to ensure that there's no contamination up front coming into your manufacturing process and this would be the step where you want to catch it because at this step the patient may still be available to get a second leukoresis and you don't want to introduce any contamination from the lucas releases process into your manufacturing process so i'm not going to read through each of these but as you can see these are high-level things that could happen at the lucas resources process and it also provides you with some mitigation actions that you can take at this step in the process to help prevent mitigate potential microbial contamination from coming into your manufacturing process and again this is one of the more critical steps because you want to catch it early any potential contamination that's coming in from either an incorrect glucoresis or from the actual patient material because at this point you want to stop it from even being manufactured and getting into your manufacturing process and i want to point out another factor is that when you're performing this microbial testing for to detect potential contamination you want to you want to have a test that can detect it quickly so the traditional companion test which requires 14 days would not be suitable in this case because you want to be able to detect the contamination quick tracks in a hurry so that you can at least let the physicians of the clinicians know that hey this patient looking for research is contaminated and we need to get another sample before we can actually begin the manufacturing process and hopefully that gives the patient a better chance of survival so at the lucas releases space these are some of the contributing factors on some of the things that you should consider considerations for a contamination control so at this point there's potential increased athleticism microbial contamination risk the production process and the patient because remember i said if you can detect the contamination before it actually gets into the manufacturing process it's better because you can then go get another april reason from the patient and start the process again and if the contaminated product does get into the manufacturing process there's an increased risk to both the production process and the patient because if a microbe is present and you then introduce it into the growth steps of the manufacturing process that micro is going to potentially proliferate which is going to then compete for the same nutrients as the patient cells and it's going to cause death of the patient cells which means it's really not good for the patient because you then have to go back and start the process again so this is the stage in the process where you want to introduce some absence of contamination testing because it provides important and valuable data because if you can detect the contamination up front you're not going to start your manufacturing process you're not going to introduce an organism to your manufacturing process that you've not seen before and typically in cartoon manufactures when we see microorganisms that come in and we get a contamination it's normally um pseudomonas salmonella's fat epi things that you would typically see of human nature the most contamination that we were seeing cartier manufacturing actually comes from the leukopheresis of the patient themselves and not from the manufacturing environment so that's one key thing to keep in mind and again you want to consider performing testing at the look for recent centers to get it up front or you either want to test it when you receive it at your manufacturing site and again because this is a very time sensitive manufacturing process both for the patient sales as well as for the patient life you want to have a rapid method in place here because you need to get your results back as soon as possible so that you can take action swiftly so let's move to the next major bucket in the manufacturing of cartier products so we have our lookout for recess material so now let's consider all the raw materials and consumables that we're going to use in the actual manufacturing process so in cartoon manufacturing we have to remember that all product contact materials are sterile they're single use and they're disposable so unlike traditional manufacturing where you have the bioreactors and the filling lines where their product actually comes into contact with those things and you need the traditional cleaning validation cleaning qualification and effective cleaning program and cartoon manufacturing everything is single-use disposable and sterile which minimizes the risk from a contamination control standpoint because if you have a robust raw material program in place where you qualify your vendors you actually qualify your materials this should be uh create a less opportunity for the ingress of adventitious microbial contamination so at this point if you have a robust raw material program in place that is another step in your contamination control for car t release and again i highlight the considerations that you should consider from a microbial contamination standpoint the raw materials and consumables actually present a decreased adventitious micro microbial contamination risk to the production process and the patient because everything is sterile it's single use and it's disposable you will prevent potential adventitious microbial contamination you would detect it upstream via your raw material program which means that it will never come into contact with your patient material so therefore this is another additional step in your microbial contamination program and in order for your raw materials program to be successful and to contain all of your um controls that you need from a microbial standpoint you need to make sure that when you bring these materials into the facility they're going to come into the warehouse and it's important to note for car cheap materials manufacturing materials that a lot of the consumables that we receive they're not triple bags so they're not the things that you will see in your typical manufacturing process so you can either get a bag and it just has one wrap on it or you can get it back and it comes directly in on a cardboard box so you have to make sure that in order for your raw material program to be effective even though they're single uses sterile and disposable remember that your production operators are going to be touching the outside of the bag so you want to make sure that when you establish your cleaning and sanitization program that you take into account the raw materials that you're bringing into the facility because you want to make sure that you physically remove all the dirt and the potential microbial organisms that are present on the exterior surfaces of those bags before they come into your manufacturing facility and this is extremely critical for currency manufacturing process because typically you're going to receive it and you're going to unpack it you're going to sanitize it and then you're going to transfer it into the manufacturing facility so you want to make sure that your cleaning procedure so your sanitization procedures are effective and robust because otherwise you're going to be transferring exactly what came in on that material into your manufacturing facility and i have a question from the audience it says um this single use is also true for allergenic therapy i think it is also true for allergenic therapy is that they also use feral signal use on disposal materials the next we're going to go into the next process in the manufacturing the next major bucket so when we say media in turkey manufacturing we're talking about all those things that we add the supplements that we add to allow for the patient cells to grow after we have made the transduction so because these are aseptic manipulations all critical aseptic manipulations in cartoon manufacturing are performed typically in a bsc which also meets the iso 5 grade 8 standards and we don't make media from scratch we simply supplement media we formulate it with sterile supplements so all the media supplements are also sterile when you're making your supplements inside of the bsc you want to make sure that you have a risk-based em program including personnel monitoring and most of the manipulations that are performed in the bfc for cartoon manufacturers are a short duration and even though all the media components that we're adding together are sterile as an additional contamination control most car t manufacturers will have an additional sterile filtration of their media so again these are all the things again that you would consider when you're thinking about your release strategy for car t products is that even when you perform activities in the grade eight area that all of these things are sterile and because you do have personnel who are making still aseptic manipulations you want to make sure that you're monitoring their gloved hands because they're the closest thing to the process that you're gonna get potential contamination from is from the person that's loved hands and i failed to mention this at the beginning of the presentation but in order to establish a process like this you need to make sure that you have done a robust microbial risk contamination assessment so you want to start from your incoming nuclear resistance all the way to your final product and you want to assess every step in your manufacturing batch record and within each of those steps you want to come up with potential contributing factors where microbial contamination may be introduced and this is going to take a while to do so you want to make sure that you have a cross-functional team assembled and all the people the smes the people who have the knowledge of the process and you want to make sure you have a microbiologist on board you want to make sure that you have somebody else from your manufacturing science and technology group with your analytical science and technology group you want to make sure you have somebody from facilities from engineering because you need to look at everything that's in place in your manufacturing area and you want to make sure that you cover every area and every step in the manufacturing block manufacturing process based on the batch record where you think potential microbial contamination can be introduced into your process as well as all those things that you need to do to mitigate those axes and let's go and look at the considerations that we'll have for media so again because all the media is sterile and we're simply formulating sterile components together and we have a terminal sterile filtration the media processing carte manufacturing also presents a decreased risk of advantageous microbial contamination and again you will have potential contamination detected upstream because all of these things are going to be a part of your raw material program you want to qualify your supplier you will have ensured that everything that comes in still is really fair and nothing with contamination will actually get into the manufacturing process so at this point you may not need to have any additional microbial testing you would simply need to make sure that your filter integrity testing passes each time you add the media supplements together and filter them so as you can see so far all of this is building on the controls that you can have in place because if you have all these controls in place during your actual manufacturing process it really eliminates the need to have that feel good testing at the end and that's why i'm leaning towards the just in case i mean just in time release or the two-day release which i'm going to get into later so now let's look at the manufacturing plot process and again i'm not going to read each of these but as you can see that there are many buckets in the manufacturing process and again in the manufacturing process every critical apex manipulation is performed within a biological safety cabinet which meets iso 5 grade a requirements most of the aseptic manipulations are short duration they are all performed in a controlled environment personnel have on the proper gowning we have an environmental margin program in place to detect any potential contamination that the personnel may have had on their fingers or their hands while they're performing any of the manufacturing and again remember that most of these are just aseptic connections that you're making and if they're the fluid pathway of the product is never exposed to the environment and that's another control in cartoon manufacturing and wherever you think that your fluid pathway may be exposed to the environment that's where you want to make sure that you're using a biological safety cabinet and you have all the environmental monitoring in there to help you to protect any potential contamination that you may be that you think may be present in your environment however if you do your microbial risk assessment and you map out every step in the manufacturing batch method you're probably come to the same conclusion that your highest risk of microbial ingress will come from personnel gloved paths because they're the ones who are making all the aseptic manipulations and their fingers are the things that will come closest to the potential fluid pathway of the product and not the actual environment so let's look at some microbial contamination considerations from the manufacturing standpoint so again the actual manufacturing process presents a decreased risk of the introduction of adventitious microbial contamination again i will emphasize that it's all sterile senior use consumables that are utilized in the manufacturing process there are no product contact services that are not sterile and that are reusable in the process for car2 manufacturing and again based on that intense and very detailed microbial contamination risk assessment that you will perform personnel gloved hands will probably come out come out as ranking the highest in your risk of potential sources of probable contamination into your manufacturing process and finally even though you shouldn't use this as a microbial contamination control strategy but it's also reality that because we do use growth media and it's doing the cell expansion that support microbial growth your process is really actually self-contamination self-contaminating checking because typically you have a minimum of five days of incubation in a nutrient-rich media that your patient cells have grown in and if a microorganism is present it's going to compete for the same nutrients in that media and it's going to proliferate as well and an example i can give of this is that one of my previous companies we had a contamination that came in with the patient however the apheresis centers did not know that they had introduced the contamination it was actually from a patient who had um they had to put the the needle in the groin of the patient and the patient had massive diarrhea but the nurses thought that they had cleansed the area sanitized and disinfected the area disinfected the area well so they assumed that there was no contamination when they sense the patient cells however three days of incubation of those patient cells the entire bed turned black and when we did the investigation it was determined that the leukophoresis process had actually introduced salmonella during the leucophoresis process and therefore the salmonella proliferated and it killed the patient cells and therefore the patient had to be re-aced again to get additional blood to start the manufacturing process and again this is one of those steps where it creates additional delay for the patient and we don't want that to happen because the patient is already at their last point of life so we really don't want anything to be introduced into the manufacturing process so if we can catch the contamination up front before the manufacturing process starts it gives greater hope for the patient because they can then get aseriesis again and we can start the manufacturing process i have two questions that are coming from the audience that i'm going to answer the first question that says how often are you doing hfa process simulations and what media issues so for after process simulations again you can take a risk-based approach it depends on how your facility is designed because people are normally the equipment for cell therapy manufacturing so your process is pretty much mimic in every suite or room that you may have so the equipment is the same the people is the same the process is the same so i think if you it would depend on your company but i think the agency is willing to accept that if you have light for like in all of your manufacturing areas and it's the same people who are performing the same manipulations on each of those suites that you only have to do one aseptic manipulations for each of those suites and you have to determine the frequency of how often you're going to repeat that accepted process simulation the media that is used for aseptic process simulations entirety is the exact same media that's used in traditional pharmaceutics triptychs or brought by any nutrient rich nutrient enhancement risk frost so the next question is is the just-in-time testing technically considered to be an in process control or a lot relief test so in my opinion the just-in-time testing is considered to be the lot release because you're saying that you have demonstrated that your process is in such a state of control that you're not going to introduce any adventitious contamination throughout the process and where you have decided to perform testing all those testing results will come back as no growth and therefore your process is in a state of control so that you don't have to perform that testing at the end of your process but again let me just re-emphasize that you have to have performed a federal microbial risk assessment you have to have going through each and every step in your manufacturing process and highlight the risk and ensure that you have mitigating accidents for all of those risks and you have to have the testing in place up front to detect any potential contamination at those points that you've seen a critical in your manufacturing process so let's continue these are some additional considerations you have established procedures in place to prevent potential cost contamination between patient lots um the potential increased risk of advertisers microcontaminations again at this point stems from the income in leukocyresis and from the patient and the greatest risk at this point would be to lay the product on no product getting back to the patient which will be detrimental for the patient and i have another question that just came in from the audience is there ever a time when the april reaches from the pace that's received is found to be contaminated but only slightly and you continue to process either you start over no matter how much contamination my suggestion would be that if you find contamination in the incoming material you should ask to get a fresh material because even if you identify the organism and you think it's a slow growing organism you never know how it's going to react when you introduce it into the growth phase of your manufacturing process and you don't want to introduce any organisms to your facility that are not typically found in your facility so that's the other concern when you actually bring in aseriesis that's contaminated from the patient you have now exposed your manufacturing facility to that organism as well even though you should have controls in place that is so that it doesn't get out in your in into your actual manufacture silly you just want to be careful that you don't put the patient at risk by continuing to process something that you know is contaminated with the potential for it to proliferate and to kill the patient cells so you the key and cartilage is that we want to give the patient as much time as the patient needs to get the drug back to the patient so that the patient can recover because every single day every single minute matters to these patients because they're literally at the point of death and we don't want to do anything that's going to delay getting that product back to the patient i also have another question from the audience does it make sense to keep the room classification that's great b area for manufacturing i am assuming that this person is asking if the bsc has to be in a grade b or a grade c area and if that's not the correct assumption the person can let me know but my recommendation would be to um based on your microbial risk assessment to determine what extra room classification you need so in currency manufacturing there is nothing exposed in the area outside the great b so everything that you transfer from your warehouse that you sanitize you transfer it into your grade d and then to your next higher classification nothing is open at that point so everything you're doing is sanitizing and passing into the bsc so in my professional opinion i think that you should qualify the room that's a great b doing your environmental performance qualification but you should operate at the grade c based on the actual manufacturing steps that are occurring in that area and if that doesn't ask you the question please let me know and i'll try to answer when you respond so next let's talk about the final product so again everything that i've talked about so far is performed inside of classified areas so the final product you're pretty much adding in your prior protectant to protect the patient cells because when you add the viral preservation material to patient cells it has the same effect on patient cells as it would have on the bacteria but if you add it too fast it will kill the patient's cells that causes death to the patient cells so there's no way to prevent that so you just have to have a validated process in place to ensure that when you're adding the chiroprotector to your cells that it's done in a way so that it doesn't kill the majority of the patient cells and the same thing will happen if what bacteria bacteria is present you add in the car preservation you want to make sure that there's not going to be any apoptosis for your bacterial cells as well so again the final product in cell theory manufacturing all the they are they consist of low risk aseptic manipulations another thing i want to point out in cartoon manufacturing is that in traditional sterile manufacturing the words open and closed they're used a lot however when you try to transfer the word open and closed over into cell therapy manufacturing there's no direct correlation because in cell therapy main factor for the most part the fluid pathway of the product is never exposed to the environment and what that means is that when you're working with the bags when you make the connections you typically clip off the two ants that you're going to make the stereo well so that your the patient blood never sees the actual environment and that's why when you do your risk assessment you're going to determine that you're the greatest risk is from the person that gloved hands because they're the ones who come closer to those aesthetic manipulations that you're making but the fluid pathway of the product is never exposed to the actual environment even though we have all the controls in place to perform them in an iso 5 grade a area all the other rooms are classified because again it's all about risk mitigation we want to make sure that we don't introduce any adventitious contamination even though the risk of introducing any advertising contamination is minimal but again you want to make sure that you're processing an aseptic manner because you want to make sure that you have taken all the all the steps that are needed to introduce controls to prevent any evidence microbial contamination so again for the final product the the risk of introduction of advantageous contamination at this point is low because you're really just making aseptic manipulations that have low risk and lastly low risk if this means that you're making sterile tubing connections and moving things from one bag to another so again i'll highlight some of the considerations from a microbial contamination standpoint again that's a decreased risk at the final product stage of introducing adventist contamination you can perform endotoxin microplasma and detection testing at this point if you like these are all the traditional safety testing sorry let me go back for a minute i just want to point out one thing so for endotoxin microplasm um remember that in cell therapy may correct and there's no water in the process so if you control your raw materials through testing up front and you don't have any way of introducing endotoxin microclassing from your actual manufacturing environment this is another thing that you should consider when you're doing your release of the product is that if you control endotype endothosin and microplasma up front from your raw material program there is a potential that you can eliminate these testing from the end and that will also contribute to your just in time release not only for the microorganism testing but also for the other safety testing and endotoxin mycoplasm but again you have to have a robust raw material program in place and you have to demonstrate that those things are controlled to the point where you know that there's not going to be any endotoxin mycoplasma introduced into your manufacturing environment so in summary i'm gonna go with the two options that i provided from the beginning that i discussed so option number one is just in time release which means that at the end of your final fill of your product and when i say final fail remember this is for cartoon manufacturing so it's not your typical ten thousand valves you would have for a traditional pharmaceutical and car t manufacturing every single patient lot every single patient that we get is considered one lot so it's one to one you get in one pace of blood that makes one lot so the justification for just-in-time release is that the patient is waiting on the product remember i said this is the last treatment option for patients this is life or death for them so we really don't want to have anything that's going to interfere with getting this product back to the patient you have the adequate engineering and procedures in place throughout the entire manufacturing process to mitigate any risk of microbial contamination and again remember i said that the highest risk of the introduction of adventist contamination and cartoon manufacturing actually comes from the lucrative process and or the patient and not from the manufacturing process the majority of the manual asap manipulations in the manufacturing process present a minimal risk of advertiser's microbial contamination and if you have all the controls in place the contamination will be readily detected upstream prior to patient needs so again if you have done your risk assessment you're performing testing in all the places that you think the contamination could be introduced you have a robust raw material program contaminant raw material program in place where you're testing all your raw materials you qualify your suppliers to qualify your vendors just in time release can be an option for car t manufacturing however if you're not willing to take the client into going to just-in-time release of carte process i am suggesting another option so remember i said that most of the all of the aseptic manipulations that i consider critical aseptic manipulations are performed inside of a biological safety cabinet so if you consider those steps that are performed inside the biological safety cabinets being your critical steps where you could introduce advocates contamination and remember i also said that based on the risk assessment that the highest risk of the introduction comes from personnel gloved hands so now if you get an id of the floor of every organism that you recover from those activities that performed in the bsc on those personnel gloves hands so you want to consider the organism that you recover in an actual bsc and you also want to consider those organisms that you recover on personnel gloved hands if you're using a rapid detection method the majority of those organisms are identified within 24 to 48 hours so if you don't want to do just in time release you can also reduce the number of dates you perform testing by only focusing on those areas where you say that microbial potential microbial contamination can be introduced into your manufacturing process i have another question that just came in if the highest risk is the introduction of the lucas cerritos sample will you wait for buy reserves follow button results on the incoming samples before starting the process so if you have a rapid method and you're focused on those organisms that are coming from a human depending on which rapid method you use you can have results in one to two days so you should wait on those results before you start the main cracking process however due to the criticality of the patient most cart companies may perform based therapy testing and they proceed with manufacturing at risk because your the assumption is that the neutral reason centers have not made any issues that the patient does not have any pre-existing microbial contamination although that can't be totally rude out but because of the criticality of trying to get results get the product back to the patient in a timely fashion you start the test but you don't wait for the results however if you start the manufacturing process and you do realize that your incoming material is contaminated and it's already in the global dma cracking process and growth is not detected in your manufacturing phase that's actually a good thing because in cartoon manufacturing it's one of the things where you can actually the doctors will actually request if you have an organism present in that patient lot to give it back to the patient because the benefits far outweigh the risk because they can give the patients antibiotics they would pretty much want you to identify the organism and if you could do a manipulatory concentration to see which antibiotics antibiotics the organism is susceptible to and the doctor will start that patient on that regime so again remember these patients are at the point of death so the doctors really want to get the product back to them it's possible even if there's a contaminant present in the planet but you have to work with the regulatory bodies but you just put that caveat out there you have to work with the regulatory body to get approval for releasing that product to the patient and typically the regulatory bodies will give you approval in those instances because again the benefits for outweigh the risk for the patient in that regards okay and i've already talked about this in my two-day release option is that if you focus on all the activities that are performed in the bsc growth is detected usually within 24 to 48 hours and the longest time for detection a typical anaerobic organism and you don't have anaerobic organisms because we don't have any anaerobic conditions now manufacturing process so you again when you're doing your method validation if you're comparing it to the compendium method you want to use the same organisms that are in the companion method but you also want to introduce organisms that are not typically in the continuing methods because in cartoon manufacturing since you're dealing with human blood you want to focus on those organisms that are going to be of clinical significance so you want to use those in the validation of your rapid method as well because those are the organisms that you're going to typically see because remember i said that most of the contamination comes in with the patient loopholes materials and they're going to be a clinical significance and not typical facility isolates and again those are all recovered typically within two days using a rapid method so here i just have some examples of some rapid methods that you can use most car t facilities use the back tee which is the one on the left and there's also the back check which you can use but most companies use the back tee because the back t offers two incubation temperatures whereas the tech only offers one incubation temperature so if you the fda has stated repeatedly that they want you to even though this is a rapid method and they know it's advancing they still want you because the companion method is still their gold standard when you do your validation they want you to compare it to compendium and if you're comparing it to the continuum method you have to have two incubations temperatures 2025 and 335 however if you're manufacturing processes that of cell therapy where all of your manufacturing blood cells are incubated at 30 35 degrees you can then solve your bla to have only one incubation temperature for your blood product because it's 335 but doing the validation in comparison to continue method you have to do both temperatures however when you do your falling you can submit the temperature where where you perform your manufacturing activities which is typically 30-35 degrees which is where you incubate yourself and that's where you expect your organization to be recovered another thing i want to be careful of is that the fda and other regulatory buyers have stated that they don't want to have a cookbook approach to telling companies what to use when you do your validations for instance how low do you go with your inoculant when you're doing your validation you as a company need to determine your own inocument level but the fda expected if you have a rapid method that it's going to be typically it's going to be more sensitive than a companion method so they want you to show that at least your rapid method can detect organisms at a higher sensitivity than that of the companion method so that's their only requirement but they will not tell you exactly which another level you have to use for evaluation i just wanted to point that out and that's the end of my presentation thank you very much and if you have any questions at the end i'll be here after laurie finishes her presentation stephen i will turn it back over to you thank you irving now before i introduce our next presenter laurie dahn once again i would like to remind our viewers that there will be a live question and answer session at the end of the webinar so please feel free to send in your questions at any time during the webinar via the questions tab located directly below your webinar screen and we'll go through these at the end of the presentation now please allow me to introduce our second speaker laurie dan is the director of scientific affairs at bio meru and has experience in clinical environmental and industrial microbiology she is a technical expert on rapid and alternative methods and participates in the sourcing and evaluation of new technologies and potential partnerships in the field of microbial control she provides scientific support to the healthcare business in north america and is responsible for managing feasibility testing and method development for biome room instruments and culture media products laurie received her phd from the university of minnesota in microbial ecology she also holds a master's degree in limnology from the u of mn and a bachelor's degree in the medical terminology from ferris state university i'll now be handing over to our second presenter welcome laurie thank you steven and hello everyone thank you irving for your informative presentation i think irving provided a good overview of car t product manufacturing and the importance of the product to the patient i would like to say that the people i've met who worked in cell and gene therapy are incredibly passionate and they know that what they do saves lives as irving stated they are many times a person's last chance of survival i've also experienced that it's not uncommon when visiting a salon gene therapy production site or or at a conference for someone to provide a personal testimonial on how these products are saving the lives of their friends relatives or even themselves so so clearly there is a need to get these life-saving products in the right condition to the patient as quickly as possible and irving outlined two strategies just in time and two-day release for car-t products using a risk-based approach in my presentation i'm going to focus on smart quality solutions for car t product release specifically focusing on automated microbial detection and mycoplasma testing uh but but before i start i would like to provide a little background on who we are uh bumari was created by dr elan mario in 1963 and he is the grandson of marcel mario who was a student of louis pasteur and marcel mary you founded the institute mario in leon france in 19 in 1897 where he developed the first anti-tetanus era so today's mar today marcel's great grandson alexander you continues the family tradition and is our current chairman and ceo this slide highlights our commitment to diagnostic microbiology burmereu provides the widest portfolio for efficient quality control solutions so these include environmental monitoring microbial identification standardized strains with our very popular bioball endotoxin testing using recombinant factor c validation services and lastly what i will be focusing this presentation on is sterility and mycoplasma testing now irving provided an overview of car t production and this slide is showing a highly simplified needle to needle supply chain cycle we know that there are numerous challenges associated with manufacturing cell and gene therapy products which i'll summarize shortly and for these products the supply chain activities must be highly synchronized and managed to successfully deliver the right product to the right patient in the right condition and again all of the testing and manufacturing for these products are to ensure patient safety i would like to highlight the differences between allergenic and autologous specifically from a manufacturing point of view the differences highlight the need for enhanced chain of custody and chain of identity and the need for smart quality solutions so for allergenic products one batch serves many patients and the product may also be cryogenically stored for weeks to months sometimes this is called a scale up however for autologous products one batch serves one patient and so this seems easy when you're managing one product or or even 10 products but what if each of these individuals were to equal 100 products how would you manage 300 products or even 700 products at a time therefore the biggest challenge that i hear when i talk to someone in cell and gene therapy is the need for chain and custody data integrity and electronic tracking now i've been at several conferences and what i've heard from the fda themselves is that they've stated many times that they focus at where there are risks for cross contamination therefore when you're performing your quality testing is important to use closed systems and test methods that provide the best data integrity so just to summarize some of the challenges of cell and gene therapy these products usually have a very low production volume and short shelf life it can be just a couple weeks or maybe only a few days there's some they're sensitive to temperature and there may be a need to maintain a cold supply chain the raw materials are complex and in fact the starting material itself is highly variable the analytical methods used for testing are highly complex and you need enhanced tracking and data integrity and you have to have the ability to scale up so the the importance of managing these challenges is again to get the right product to the right patient in in the right condition so coming back to our cell therapy production process an additional challenge to quality testing is that it is a continuous process and it takes days not months so a few examples from a recent conference showed a typical production cycle of 14 to 20 days so the goal again is to get the cells back into the patient as soon as possible and these products cannot be terminally sterilized and the process following aseptic techniques so there is no time for end product testing following traditional methods but rather a risk-based approach is used similar to what irving ford described in the previous presentation therefore a robust quality management system must be established using smart quality solutions an additional challenge is that the majority of testing is performed in a separate centralized qc laboratory there's a lot of value in finding opportunities for rapid testing and bringing this testing out of the lab and directly to the production floor and the best way for this to happen is to work with suppliers that offer rapid solutions now in my presentation i'm going to focus on two rapid testing solutions that are ideal for cell and gene therapy and the important thing is that they are simple to use fast and accurate in irvine's presentation he highlighted the use of atmp guidelines and here i describe the three main types of rapid microbial methods that can decrease the time to result from the 14-day compendial test that's described in both the ep and usp now alternative methods have been embraced and they're described in ep 516 and usp 1223 and they include growth-based methods such as co2 production and bioluminescence direct measurement such as solid phase cytometry and flow cytometry as well as cellular component analysis more recently the ep has published 2627 to address microbiological examination of cell-based preparations that also includes the use of rapid methods there's also a new chapter usp 1071 that was recently published in june of 2019 and will become official in december of 2019 for rampant testing of short short life products using a risk-based approach so what i've seen uh really in the past five to ten years is that there's really been a shift from rapid methods being an alternative to the compendium test to the compendium test being a rapid method so rampant methods can reduce the time to detection from 14 days to as little as four hours so it's one growth-based rapid method technology based on co2 production that is ideal for cell and gene therapy products is the back t-alert 3d microbial detection system the instrument is modular for example here is the batch alert 3d combo so it combines all of the backseat alert 3d capabilities in a compact format it includes a control and incubation module in a single instrument and it has a total incubation capacity of a hundred and twenty cells here is shown the battery alert the 3d 240 system that consists of a control module on the left and one incubation module on the right that is made up of four incubation drawers of 60 cells giving a total capacity of 240 cells so the control module can actually enable up to six incubation modules so providing that flexibility so therefore the full instrument configuration can allow incubation of 1440 bottles now as irving mentioned in his last presentation the flexibility and modularity continues with the battery alert 3d dual t system now this configuration has both a low and high temperature incubation modules allowing it to closely follow the pharmacopoeia guidelines so the low temperature module allows for incubation at 20 to 25 degrees and the high incubation module allows incubation at 30 to 35 degrees so similar to the single temperature modules additional incubation modules can be added now with the batch alert system the only reagents that are needed are the aerobic and anaerobic bottles that have unique barcodes so once the sample is added to the bottle you have full data integrity the batch alert system utilizes a color metric sensor and reflected light to monitor the presence and production of co2 dissolved in the culture medium so once models are loaded the color metric sensors are scanned every 10 minutes microorganisms produce co2 as we metabolize the substrates in the culture media and this changes the color of the gas permeable sensor in the bottom of each culture bottle from blue green to yellow so once growth is detected this the system alarms both audibly and visually and the sample data is recorded now the testing of the batch alert system is incredibly easy it's just three steps you inoculate the media bottle then you scan the bottle and then you load them into the incubator i've talked to a lot of cell therapy companies and this is what they love about the system they love that it is so easy so there's no worry about removing enriched sample or performing additional manipulations or testing that may include reagent management so again where there's a high need for chain of custody and chain of identity and data integrity you want to use a smart quality system like the batch alert 3d now the batching alert system is an automated culture method the bottles are continuously monitored reading the bottle sensor every 10 minutes the continuous monitoring allows for faster detection of positive results and the reading is objective and completely independent of turbidity so there's no need for subculture the dual piece system allows for incubation following the harmonized pharmacopoeia guidelines and is 21 cfr compliance now there have been numerous publications using the vascular system for traditional pharma as well as cell and gene therapy product testing and recently the nih published in the whoops let me go back one i'm sorry recently the nih published in the journal of clinical micro in february of 2019 comparing the traditional and automated culture systems on 118 microorganisms this is probably the most comprehensive study that i've seen and the results showed that the bachelor system can be used as an alternate sterility test now with that said most cell and gene therapy companies using a risk-based approach are focusing on clinically significant microorganisms that are usually detected within two to three days and as irving stated um their contamination is being detected within 24 to 48 hours so now i'd like to shift to our second rapid microbial method and that is microplasma so just to give everyone an overview of the challenges of this microorganism it is ubiquitous the cell cultures provide a very favorable environment for their growth and if you have a contamination event the quality and quantity of the product is decreased and some of the species are clinically significant mycoplasma is difficult to prevent it's difficult to detect and it's very difficult to eliminate if you have a contamination in your manufacturing facility so especially for cell and gene therapy there is a need for a fast simple and robust solution so in cell in gene therapy the traditional method really can't be formed cannot be performed due to the long testing time this slide shows the traditional method that can take 28 days for results here's presented the indicator cell culture method and while it's faster than the culture method it still takes five to eight days and requires a lot of technical expertise now there are alternative molecular tests and i think most cell and gene therapy companies are using molecular test systems they're available to provide results in about five hours but the test method requires expertise and many times a molecular biologist is needed the test is prone to contamination due to the open test system and a specialized laboratory is required so at this point i mean what if i could tell you that you could take a 28-day traditional test or even a five-hour nucleic acid test that requires a high level of expertise and turn it into an hour and make it so simple that anyone can perform it it doesn't require a special lab and it could be potentially performed directly on the production floor so what i would like to introduce is the molecular lab in a pouch for testing mycoplasma this is the biofire mycoplasma assay and it's the fastest easiest and simplest mycoplasma test method available the test is completely closed system for performing molecular mycoplasma testing and takes less than two minutes of hands-on time and provides results in about an hour it's so simple it can be formed it can be performed by anyone anywhere and at any time the bio fire system consists of two components the film array instrument and the single use molecular lab and a pouch disposable so just with these two items you have everything that you need to perform fast accurate microplasma testing so the the compact film array instrument performs the extraction amplification and detection in a single machine you can actually connect up to eight instruments together for higher throughput now we've completely transformed the microplasma analysis from something very complex to something very very simple so this pouch contains everything to realize a molecular analysis from sample to result so the sample is injected and goes from the lysis chamber to detection chips and includes 14 pcr assays designed to detect more than 120 mycoplasma species it contains all the controls necessary to control each step of the reaction the reagent pouch system also decreases waste because one pouch equals one test and it simplifies inventory and that the pouch itself is room temperature stable so just giving an overview of the steps similar to the back alert uh that it's something very simple to do uh the the the workflow for the biofire mycoplasma assay is also simple you simply inject the hydration solution and unprocessed sample into the pouch you place the pow pouch into the instrument and start the run and the software reports whether or not the microplasma is detected all this happens in less than an hour you have easy to interpret results the report that's generated shows that you either have mycoplasma detected or mycoplasma not detected so again comparing the different methods the traditional method for selling gene therapy is really not feasible at all to perform since it takes 28 days so just looking at other nucleic acid methods uh compared to the biofire method you can see that the biopharma method is shows a significant reduction in time a novice can perform the the assay you have low contamination risk because it is a closed system and the reagents can be stored room temperature and can be performed anywhere so just to summarize the new biopharmicoplasma assay the test is easy again there's no pcr skills that are needed there's no special lab that's required no precise measuring or pipetting you have very minimal data entry it's automated and it provides simple standardized results it's easy it only requires two minutes of hands-on time and the sample to result in less than 60 minutes and finally it's comprehensive it can detect more than 120 strains of mycoplasma and molecuities you can test raw material in process and final product it's 21 cfr part 11 software we provide full validation support so the benefits of implementing a rapid contamination test such as the bachelor and the new biopharmicoplasma includes the use of automation and improved work efficiency and data integrity but it also provides standardization ease of use and objectivity so both solutions are easy and they can be brought out of the lab and close to manufacturing where they are needed so to summarize cell and gene therapy production is highly complex with numerous challenges there's a need for enhanced chain of custody and data integrity to ensure the right product is delivered to the right patient without cross-contamination new regulations allow risk-based a risk-based approach for microbial contamination and microplasma testing and new alternative methods are moving the lab to the production floor so with that i would just like to thank brian whitehouse and marianne chu for providing some of my content today this is my contact information if any of you have questions about the bachelor or the microplasma assay with that i will hand this over back to stephen thank you thank you laurie for that great presentation uh now we will begin the question and answer segment of the webinar for once again i would like to remind the audience that you can still send your questions in via the questions tab located directly below your webinar screen so now our first question will be for irving and our first question is what is the biggest manufacturing challenge for car t products um the biggest challenge is hiring personnel especially in the area where we are in new jersey um there are about five car t manufacturing companies here so we're all competing for the same resources and that's one of the biggest challenges the other challenge is that uh because one patient does equal one lot and there's lots of variability and that patient sells um it's it's very difficult to know if a patient not they don't act the same so when you're manufacturing there's just a lot of variation in the process so it's never a standardized process so i always tell people who are going to work in cell therapy to forget your typical manufacturing life and you have to come into a mindset because everything in cell therapy is great it's no longer black or white and you have to be able to make decisions instantaneously on the floor and investigations can't take the typical 30 days you have to have a triage right away because the patient is waiting and the cells are really time sensitive so you have to have a mindset for your quick your quick pick or on your feet because you have to make a decision live with that decision and keep the process moving thank you our next question is are the regulatory bodies on board with such risk-based approaches leave that again it's for you irving yes that is for me so i i was just um at the pda global micro conference and i was speaking to some of the reviewers and lead investigators and they actually um asked me for my presentation because they said they actually like the fact that companies are now taking a holistic and comprehensive approach and actually mapping out the process from a risk-based standpoint so they're they're actually on board with it i don't know if they are ready to jump into the whole just-in-time release but they're definitely on board with the reduced um timeline at the end but i think if more cell therapy companies come on board and the more important it is to get the product back to the patient in a hurry and the more that this becomes the standard treatment for people instead of the final treatment that the regulatory bodies are going to be more accepting i think it's a matter of the company defending their process or their staff and making sure that they have the data and the risk assessments to support it thank you our next question is with the backed alert system does bio meru have media available that allows neutralization of antibiotics and that'll be for you lori oh yes yeah good question um the media that we have available we have standard bottles for both aerobic and anaerobic and we also do provide resin bottles and the resin bottles are designed to specifically neutralize antibiotics so the answer is yes we do have media specifically for neutralization of antibiotics thank you and our next question is when will the bio fire microplasma test be commercially available that'd be again for you lori oh yeah um currently the the test is in pre-market customer testing we anticipate the launch uh for commercial availability in the us in march of 2020.

[01:12:07]fantastic um moving on our next question is how important are rapid methods to car t manufacturing and that'll be a few irving oh they're extremely important actually the regulatory bodies um that's one of their key questions when we're doing inspection of car key facilities is how can you prove to us that based on the number of patients that you're going to get that you can actually manufacture them and get them back to the patient in a timely manner and as a result of that question they do expect you to have rapid methods because the traditional um 28-day microclass of 14-day stability they do not fit into the whole car-t platform because the manufacturing process for car t products is typically only 10 days so you can imagine if you have a process that the patient is buying and they're waiting for 10 days to get their product you don't want to delay it another 14 days for waiting on the results that's really not going to provide value so they they really expect you to have record methods when you file an application for sale therapy product thank you our next question is um for lori our next question is does the bio fire microplasma identify the species of microplasma um yeah we get this question quite a bit the the answer is actually designed as a presence absence test so it doesn't identify the specific microplasma species but rather microplasma detected or mycoplasma not detected thank you and with that we will actually conclude the questions uh and answer session but thank you all members oh steven i had a question that came in that i don't think we gave an answer to i just want to make sure that we answered it what what what is an acceptable endotoxin specification for cell therapy products this could express that eu per mil or eu per kilogram per hour it is actually expressed at eu per ml and you would typically use your um average body weight of the patient population to determine the endotoxin calculation that you would typically do for standard pharmaceutical products basically i'm sorry for missing that question great and with that we will conclude the question and answer session uh before we finish the webinar um i'd like to ask our presenters if you have any closing remarks for our audience um so irving is there anything you'd like to let our audience know about yeah i would just like to thank the audience for actually participating today in the webinar and party is an exciting world and if you're not working in cartier i would greatly encourage you to get into the cars you feel because i believe that personalized medicine is going to be the way of the future and car t manufacturing is very never a dull moment or a boring moment and i i i'm really proud to say that i work in a manufacturing environment where i actually can see the results going back to the patient and i'm actually saving patient life and i can actually see the results of changing people's lives so i'm very proud to work in currency and again thank everyone for following me today thank you and laurie do you have any closing remarks for our audience today yeah i agree with irving thank you for your comment um it is it's a very exciting area and i'm very interested to see how things will be evolving uh with the guidelines especially uh for the risk-based approach um the just-in-time in the two-day release i i think makes sense providing you know risk risk risk that's the that's the big word to use for for car t production is a risk-based approach great and i would like to take the opportunity to thank our presenters once again that was irving ford head of the car tqc laboratories at celgene and laurie dumb farmer microbiology scientific director at biomeru thank you for sharing your knowledge with us i would also like to remind our audience that you can view this webinar and other webinars on demand by visiting biopharma hyphen asia forward slash ondemand hyphen webinars and if you are watching this on demand then please feel free to send your questions over to me at s-t-e-p-h-e-n-dot edwards at biopharma hyphen asia.com and we'll get those answers send back to you thank you everyone have a great day