0:00:01 Hi everyone. My name is Jessica Rauchet and I am the laboratory manager and senior research associate
0:00:07 at VitraVax. I just want to thank forge for the invitation to the summit. I am just so happy to be
0:00:15 included with such an amazing group of scientists. Um So today I’m going to be talking about atomic
0:00:20 layering for pharmaceutical applications, where I will be specifically talking about thermo stable,
0:00:26 time released, single shot vaccines. I know this is a virtual event, so feel free to write down your
0:00:32 questions as I go and reach out to me after the presentation. Um so just to give you a little bit of
0:00:38 background about myself, I’m originally from pennsylvania, born and raised. Um I earned my Bachelor
0:00:45 of Science and biotechnology with a minor in biochemistry from Shippensburg University and then I
0:00:50 went on to earn my master’s and biotechnology from johns Hopkins University. I have a background in
0:00:56 infectious diseases, translational cancer research, personalized medicine, asset development and the
0:01:02 vaccine development. I also worked across various sectors in government, academia and industry. Um
0:01:08 But enough about me, I’d like to spend this talk discussing the Vitra vex technology platform
0:01:13 creating thermo stable time released single shot vaccines. So if we go on to the next slide um So
0:01:23 Vitra Vac uses spray dried formulation um consisting of antigens and achievements along with disa
0:01:29 Carides. This combination allows us to obtain a higher glass transition temperature for the
0:01:36 particles as well as former glassy matrix to provide thermo stable micro particles. Um So after
0:01:43 spray drying using atomic layer deposition, we can coat these and again regiment containing micro
0:01:48 particles with a protective metal oxide. Um To provide times released doses, these doses allow for a
0:01:57 boost up to six months after injection, eliminating the need for multiple shots. Basically the L. D.
0:02:04 Layer provides a slow time the release of the particles, but also allows for extra event when using
0:02:10 the aluminum wire.
0:02:15 So if we just kind of dive a little bit deeper into spray drying. I just want to do an overview of
0:02:21 the process for those of you who are unfamiliar with how it works. So we start with the antigens and
0:02:27 the achievement along with the uh sugar formulation. This will form a glass matrix. One spray dried.
0:02:35 The formulation mixture is loaded into a syringe and a syringe pump is used to deliver the
0:02:41 formulation in a needle with heated gas. Here the formulation is atomized by the needle actually
0:02:49 pressing down and dispersing the particle mixture or the mixture itself into particles. So um
0:02:58 basically um it’s exposed to hot air which allows for rapid drying of the droplets in the drying
0:03:05 chamber and then the dry particles can then go on through the cyclone and be collected into a vial.
0:03:11 Um This powder can then farther be dried in preparation for atomic layer deposition.
0:03:22 So once these particles are spray dried, we can actually look at these under a scanning electron
0:03:27 microscopy uh otherwise known as ECM. Um As you can see, there is some variation in these particles
0:03:34 uh but also due to the components in the formulation, the particles can become wrinkled or have
0:03:40 dented appearance. Uh flow cam um is also used to help us determine uh the particle size
0:03:50 distribution of the spray dried particles. Um As you can see a lot of these do fall in the 4-8 range
0:03:59 here. Um And even looking at this ECM, you can you can definitely see these distribution of the of
0:04:08 the sizes. So um obviously when with spray drying comes some challenges as you could see from before.
0:04:17 So, due to the size distribution caused by spray drying, um we take an average particle size of 5-7
0:04:24 microns. As this helps in later applications when we go to coat for atomic layer deposition. Another
0:04:32 challenge comes from the irregular shapes and the surface area of the particles. The wrinkled shape
0:04:37 actually creates higher surface area, so this needs to be accounted for when calculating the amount
0:04:42 of precursor needed for dozing. Um And lastly, the water content in the spray dried particles can
0:04:49 also cause unwanted reactions with the precursors during atomic loitering. So after spray drying,
0:04:55 the particles have about a 5% moisture content. If we were to keep the moisture content at 5%,,
0:05:02 these particles would gain significantly more eliminate due to the reaction between try method,
0:05:07 aluminum and water. Therefore, we further dry down these particles under vacuum to decrease the
0:05:13 moisture content to less than 1%. So um we actually measure this by Karl Fischer titillation and we
0:05:20 just always try to Make sure that we’re under that 1% to prevent that extra wiring that we are not
0:05:27 accounting for.
0:05:34 So um for the atomic layering side of things. I’m not really going to go into a lot of detail
0:05:43 because I know um this is an L. D. Summit so I’m sure you’re going to be hearing a lot about that
0:05:50 process. Um So I’m going to be kind of talking a little bit more about what we do with this process.
0:05:56 So um basically as I said before we use time ethel try meth, aluminum and water as are precursors
0:06:04 also known as T. M. A. Um So due to the high vapor pressure of both the reactant, we can actually
0:06:10 run r A. L. D. Cycles at lower temperatures. This is important when thinking about bio molecule
0:06:17 stability. Another interesting feature about our process is we usually run between 50 and 1000
0:06:24 cycles. So we have a pretty wide range um And we conduct this process in both the fluid eyes bed
0:06:32 reactor and a rotary reactor. Um We don’t use both at the same time but we do have processes for
0:06:39 both. A fluid eyes bed and a rotary. Um The rotary reactor is nice though because we are able to
0:06:46 scale up for an Upwards of 100,000 vaccine doses. So that’s really awesome.
0:06:55 So to give you a little bit more about the fluid eyes bed, uh nitrogen is actually flowing through
0:07:01 the bottom of the system along with the precursors. The flow can be adjusted until the particles are
0:07:07 actually fluid. Ized and then suspended in the chamber. A vibration and or an impactor can also be
0:07:15 used to keep these particles from sticking to the walls of the reactor and everything is done under
0:07:21 vacuums to the precursors can be delivered at low temperatures to accommodate the androgen and
0:07:27 achievement embedded particles. Um The biggest issue though with the fluid ice bed um is what is
0:07:35 considered the Brazilian nut effect, basically due to the size distribution of the particles. When
0:07:42 the powders are agitated in the reactor, it actually causes uh the larger particles to float and
0:07:49 congregate to the top while the smaller particles fall to the bottom. Um This can cause uneven
0:07:54 coding if you are not being careful and conscious while actually coding the powders. Um This is
0:07:59 something that we have seen and we can adjust for that, but it has become an issue before. So we
0:08:07 started moving over to the rotary reactor. So to give you an idea a little bit more about what is
0:08:13 nice about the rotary actor is it allows for rolling of the particles inside the reactor um allowing
0:08:20 for an actual even coding. So um as you can see there is a larger chamber that doesn’t move around
0:08:28 um and then a smaller chamber inside of it, which is actually the reactor chamber that holds the
0:08:33 powder itself. So um this smaller reactor chamber is able to just roll around and that’s what
0:08:43 determines how the powder is actually rolling. Um So we can adjust that speed and direction of it
0:08:51 depending on how we actually want to manipulate the powders themselves. So the idea is that we are
0:09:01 more trying to target a rolling or cascading um kind of effect where we are constantly seeing a
0:09:09 tumble of the sand particles. Um things we want to avoid is something like this over here in the
0:09:16 corner where we’re spinning too fast and the powders are just kind of being stuck to the walls. Um
0:09:23 What else? What also is nice is, you know, we can add in other different vibrations and impact ear’s
0:09:31 to also prevent sticking to the walls. Some powders that we do use can be a little stickier than
0:09:38 wanted, so we can tweak the rotation based on the powder properties.
0:09:47 So just another view of the reactor, this is showing the precursors off to the right there kept out
0:09:52 of the oven to prevent overheating um and then they can be delivered into this bumper chamber here.
0:09:58 So the smaller chamber front here is called a bumper chamber where it allows us to actually dose are
0:10:05 precursors based on a pressure. So um it allows for a lot more consistency when it comes to dosing
0:10:13 and it makes the calculations a little bit easier with the amount that we need to do. Um Because our
0:10:20 temperatures are lower than normal L. D. Reactions. We do increase our purge times to prevent that
0:10:28 CVD build up.
0:10:32 So just to kind of do a little overview of the actual Pandora system. Uh So the writer reactor runs
0:10:40 on the Pandora software system here. I can create recipes and programs and see live graphs for the
0:10:46 temperature and pressures. Um And I can also control all the valves manually. The green shows open
0:10:52 valves and the gray are closed. Mhm. Uh You can control the speed of the rotation and um if you want
0:11:01 you can change the direction like I said before so you can um continue clock with wise and then
0:11:07 maybe do a half turn counterclockwise um and just kind of play around with that, making sure your
0:11:13 powders are not sticking to that actual reactor bed. Um So another thing that’s really nice, there’s
0:11:21 two places that read the temperature um and the same for the pressure. The temperature allows you to
0:11:26 see what the reactor bed is actually at and the oven itself. Um Whereas the pressure gauges show the
0:11:32 pressure in the bumper chamber as well as the reactor chamber. So this is what allows us to dose
0:11:37 based on pressure. So we’re able to dose that smaller chamber and then because of the pressure
0:11:42 difference between the smaller bumper chamber and the actual larger reactor were able to deliver
0:11:47 straight into the reactor for it to react with our actual powder product. Um Again this makes for
0:11:55 very nice things. Another interesting feature is these weight conditions so we can um either do
0:12:02 different weight conditions based on pressures that we’re waiting for, temperatures that we’re
0:12:07 waiting for um and other various features uh which makes it really nice. So if we look at the ECM or
0:12:19 the skinny electron microscopy of the atomic layer, deposition coded spray dried particles, we see
0:12:26 that these particles become a little bit more around. You don’t see as much of the dense, so they’re
0:12:31 much smoother. The flow ability is much better of these particles. Um this specific one over here
0:12:37 has about 250 coats on it. Um And then on the right, you can see our prediction or expected. We
0:12:45 gains based on the amount of aluminum layers that we put on our cycles that we put on. So based on
0:12:54 Calcium nation, we can actually, so calculation heats up the particles to around 950°C to burn off
0:13:02 any salts and organic so that we can assume the weight left is just our aluminum. So we can back
0:13:08 calculate um and determine if the particles received the expected weight gain. So based on the
0:13:15 number of cycles we can expect a certain weight gain and we determine it from there. Um So we have a
0:13:22 nice kind of graft to go off of
0:13:30 another. Look at our particles. We can look under transmission electron microscopy, which is TM.
0:13:36 This allows us to look at the coded particles and see the contrast between the spray dried particle
0:13:41 and the actual aluminum coding. So you can see in this photo here there’s actually a color charge
0:13:48 difference between the particle itself and the actual wiring that’s going on. And then if you look
0:13:55 over on the right here, we can see. We actually used a Fib ECM, which is a focused ion beam, um
0:14:06 scanning electron microscopy. So with this focused ion beam were actually able to cut through the
0:14:12 particles themselves and see the actual layering going on. So um after we cut through, we uh we can
0:14:20 do ECM and then we can see these aluminum wires and actually measure how much aluminum is expected
0:14:27 to be on the outside of the particle. Um so I guess the big question here is what are we actually
0:14:37 doing with these coded and again and adjuvant containing particles?
0:14:45 So the biggest experiments that we’re doing are actually time to release vaccines in mice. So how do
0:14:56 we do that? Basically? We go through all of our steps. We take our formulation with the A given and
0:15:01 an injun containing formulation. We make that into a spray dried powder and then we’re able to coat
0:15:07 that powder. So based on the number of coatings, we can expect a certain release by a certain time.
0:15:14 So using different mixtures of uncoated and coated particles, we can create boost effects so that we
0:15:24 don’t have to do more than one shot. So the idea is that we can get one shot and it’s still getting
0:15:31 those boost effects two weeks later, six weeks later, eight weeks later due to these liar ng using
0:15:40 atomic layer deposition. So it’s really cool that we are able to hopefully eliminate these multiple
0:15:49 shot vaccines and get it down. Get all these vaccines down to a single shot. Um but it’s exciting
0:15:58 new field and I’m excited to be a part of it. So if anyone has any specific questions, feel free to
0:16:06 reach out to me. Um just to kind of quickly go over kind of what the study showed last year, if
0:16:13 anyone has seen, um we published a paper using HPV. Getting it down to a single shop. So in mice we
0:16:22 actually injected them with either coated or uncoated Particles or just their regular vaccine
0:16:30 themselves. And then over the course of 14 weeks we measured the intent or the fluorescence of the
0:16:38 HPV inside of the mice themselves. So we were able to see throughout um using antibodies that we
0:16:46 were able to see um expression. Um 14 weeks out showing that we’re still getting release of these
0:16:55 particles and of these antigens into the body. Um so now we get to see the boost effects of
0:17:03 everything. Um so again, uh questions or comments. Feel free to reach out to me. My name is Jessica
0:17:10 Rocket. Again, I work for Vitra vex. Uh Thanks again to forge nano for inviting me to come speak. Uh
0:17:19 and I hope everyone enjoys the summit. Thanks bye.