Hi, everybody. Thanks for coming out this week to teach us about all of your AUC things.

It's been so great and I'm really happy to the organizers for organizing this and for

everybody for traveling as far as we did. Okay. This is my disclaimer and intellectual property

notice. Okay. So we all kind of know what the analytical ultra centrifuge is because

we're here at this conference. This image here is of the Optima AUC which we actually

brought all the way here and have it like functioning. If we wanted to do a run today,

we could. But I don't know if we have any cells, so probably not. But yeah, it is our

newest instrument. It does have the multi-wavelength capabilities which is exciting. If you guys

want to know more about it, stop by the booth. As well, when you're at the booth, you can

fill out the little form to win a brick model that you can build your own centrifuge that's

little from. So my talk today is on lipid nanoparticles. Lipid nanoparticles are being

used in the drug delivery field. They're also being looked at for gene therapeutics and

have been used to deliver fluorophores and stuff. They are composed of lipids which then

encapsulate the drug. This drug can be anything from nucleic acids like RNA, DNA, to small

molecules like we saw with the doxorubicin for cancer treatments. And then it could also

potentially package proteins and peptides as well. So the LNP is made up of a lipid

exterior shell that has carbohydrates embedded in it to help with the stability of the nanoparticle.

And then it also has pegulated lipids that help it within circulation and help to prevent

it from degradation when it's injected into the bloodstream. There are a couple different

formations that lipid nanoparticles can have. This one has the exterior shell and then

a few little my cells inside of it that hold the drug. You can also have bilayer as well

as solid core lipid nanoparticles. So there are a few different options. But yeah.

So to start with, we're actually going to start looking at RNA characterization with

the AUC to show that you are able to use the AUC to characterize your final drug product,

but you can also use it throughout the process to look at your drug or your API beforehand.

So this study was some of the work that I did in collaboration with Dr. Truchard Patel's

lab at the University of Lethbridge. I guess Patel is down here. So they were looking at

a single-stranded RNA virus. This is just a schematic of what its genome looks like.

However, in order for it to replicate, it needs to cyclize. And that cyclization occurs

by an interaction at the five prime terminal region and the three prime terminal region.

And the idea is that they wanted to look at this interaction and see if they could figure

out which nucleotides was kind of causing that dimerization of the particle so that

theoretically, if you could find that interaction, you could create a drug to disrupt it and

therefore prevent replication of your virus. So they selected a 225 nucleotide region from

the five prime terminal region and a 225 nucleotide region from the three prime end.

They have different sequences with the same number of nucleotides. It's kind of funny

that they picked the same number of nucleotides because it ended up making experiments a little

bit hard because they, on SEC, come out at the same volume. SecMALs, they have the same

molecular weight. And they also found when they ran this over, SecMALs, that they always

had dimer formation along with their monomer. And they did a lot of purification and couldn't

get rid of the dimer, so it was just an equilibrium that was always occurring. But because of

this, when they mixed the five prime and the three prime together, they would see that

dimer forming, but you couldn't be certain that it was a five prime, three prime dimer

that was forming and not like a increase of three prime dimer on its own forming. So that's

when they came to AUC. So here we took their three prime RNA and we put it in the AUC and

we got our major peak here at about 5.3S. There's a few other peaks. The one close to

seven corresponds to what the dimer would be. And then we had these two additional peaks

that we weren't totally certain of, but they were low, so we continued on. And we found

that the five prime terminal region, also 225 nucleotides, had a different sedimentation

coefficient. And it sedimented up here at about 5.9S. And the reason for this is that