Rapid Timeline to Tox Starting with KBI Biopharma's SUREtechnology Platform™, Powered by Selexis®

Hi, everyone. I'm Sigma Mostafa. I'm the Chief Scientific Officer for KBI Biopharma.

And, in my role, I am responsible for cell line development, process development, and analytical and formulation sciences groups. And I'm really excited to talk about our newest offering for cell lines, from our Selexis platform.

So a little bit about KBI. KBI is a top ten CDMO.

We offer mammalian services, from our North Carolina and Geneva, Switzerland sites and, our microbial services from our Colorado site. And then we also have analytical services as a standalone offering.

And we have a deep expertise as you see the numbers, as well as, long, you know, track record of delivering on the promises we make to the customers.

So a little bit about Selexis.

Selexis was founded in 2001 and was then acquired by JSR, our parent company, in 2017.

And subsequently, in 2023, it became a single organization with KBI.

And, one of the things I want to point out is even before twenty seventeen, there were substantial number of programs that, were coming from Selexis into KBI, into my group. And one of the things we understood from the get go is that the Selexis cell line is, particularly robust and high producing compared to the 25+ CHO platforms that we were working with from across the board from various vendors.

So Selexis has a very large, portfolio of technologies and new solutions. I'm just going to focus on a couple of them. One is the Selexis Genetic Element, that was developed in 2002. This is our primary driver for the high productivity.

Also, the CHO-M cell line development, which is our host cell line. And this is a cell line that has a very fast doubling time, fifteen to seventeen hours compared to what's in industry standard, maybe twenty two to twenty four hours. And that itself gives a big advantage to the cell line. Also, the, SURE CHO-M+ libraries, and these are for specific challenges that we might have with the protein. And for expression of the protein, we can go to these host libraries.

And then one of the things in the pipeline is the first drug in the market with the Selexis line was in the 2018 time frame. Now we are actually, we have even crossed the tenth drug. Yesterday, I found out there's eleventh drug in the market with this cell line.

So between 2023 and 2024 we, have worked on a couple of new technologies, and I'm really excited to talk about those and introduce those in this conference. One of them is our transient transfection platform, and that is using the exact same CHO line, CHO-M line, but in a transient transfection mode. So you are getting essentially the same host cell, same product quality, but, you are getting the fast turnaround and getting the material. And then I'm gonna talk about the transposase line, transposase-based technology for our semi-targeted integration that's leading to, a substantially more consistent as well a high productivity cell line.

I'm also gonna talk about our afucosylated knockout cell line. So that is for afucosylated proteins as well as, a little bit about our NGS sequence, identity, some of the offerings there. One of the things I'll mention is the basal media here is one that KBI developed. It's, marketed by Ajinomoto, and we get, very high productivity, with, the Selexis line as well as other CHO cell lines using this media.

So some of the numbers, there's, now, upward of ten eleven marketed products using this cell line as well as, 175 drug candidates that are, using the Selexis platform.

So in terms of the the overall how the CLD group is set up, we have cell line engineering, cell line generation, cell line automation, and cell line lead characterization.

I'm not going to talk about the cell line automation portion today, but we have done quite a bit of innovation there and have been able to essentially make it hands-free automated platform for cell line passaging.

In terms of the SUREtechnology, powered by Selexis, it's not just for mAb, but for bispecifics, Fc-fusions, antibody drug conjugates, recombinant proteins, enzymes, glycoprotein vaccines, and biosimilars, we equally are successful.

And, the main words and main characteristics of the cell line that I would point out are that it's rapid because of the fast doubling time, stable cell line, cost effective, and robust.

This slide essentially just points out the varied scaffolds and structures that we have already worked on. And that's where Selexis' main core expertise and their their sort of value proposition comes in because of the, you know, likelihood of us being successful with complex proteins, just based on our experience level and the wide range of problem solving we have done.

So the workflow starts with, of course, the lead protein and doing, essentially, leader peptide selection as well as, doing codon optimization followed by, Selexis Genetic Element-based, this vector development as well as, subsequent to that, the transposase semi-targeted platform for our transfection.

And then subsequent to that, either BEACON or ClonePix is used for single cell cloning. And we would do typically Ambr-based studies for all the productivity tighter phenotypic studies, stability studies, and then a high throughput early stage product quality assessment from from the get go. And the package that comes at the back end of it is highly comprehensive as well as, obviously, in terms of the output that you are getting is going to be, you know, world-class.

So a little bit about, the SGE elements. It's essentially an element that allows for, more the DNA becomes more rigid, and it allows for favorable conditions for gene of interest expression.

And some details around the SGE elements here, these are based on dA-dT sequence.

These are transcription factor binding sites, enriched in transcription factor binding sites, usually, nucleosome-free gaps. So it allows for RNA polymerase two to sit there as and allows for better transcription, which is the reason for high productivity.

And this slide essentially shows, with SGE the higher productivity we are able to achieve and and the reasoning for that.

The next thing is the host cell line. It comes from a CHO-K1 cell line.

And, essentially, the host cell, was chosen, the derived CHO-K1 cell line for its robustness as well as its doubling time. So, some of the things to point out here is that we have done whole genome sequencing.

And, therefore, we we are able to do a lot of deep analysis around cell line, including monoclonality and analysis using next-gen sequencing.

The other thing phenotype was, as I had mentioned, fifteen to seventeen-hour doubling time and consistent high productivity.

These are the other libraries we have, and the libraries are to solve specific problems, either around cell growth, metabolism, ER secretion, glycosylation, trafficking of the product, ER folding. And there's a whole range of libraries in, with chaperones and such to solve, you know, specific challenges with the protein production and therefore create bespoke solution for your product.

In terms of some of our genomic characterization, monoclonality assessment, we have, bioinformatics tools. We have a next-gen sequencing AI group that's part of KBI. And that group brings in a couple of these, solutions that not of not all cell line developers typically have, which includes single nucleotide variants analysis as well as looking at monoclonality assessment through assessment of the overall sequence and the genomic characterization in terms of transgene. genome junction prediction, and such.

This slide is just to give a picture highlight that the activities that happen after CLD are happening in a staggered manner. So, they're not all sequential.

They have to be staggered for timeline purposes.

And, what we are able to do is because of the consistency of the product, and and the cell line. We can do a lot of upstream and downstream and analytical work with early-stage material and be able to move the process through for FIH very fast. And I'll talk about the timelines we are proposing with this new offering.

So in terms of some of the timelines for the transient offering, in two weeks we we can have the initial material. Whereas, typically, when we are looking at variant selection, it's a lengthier process because we are going through multiple steps. There are pauses as well, for assessment, etcetera.

Our SUREmAb offering of nine weeks is for standard mAbs, from trans, from transfection to stability study. And one of the things we are saying with the semi targeted transposase-based solution is we can decouple the stability study from critical path and can move even faster.

So the original offering, from, this SUREtechnology, Selexis platform, is obviously we have the SGE elements. We have the CHO-M cell line with the very fast doubling time. And we have all the, cutting edge technology in terms of single cell cloning as well as high throughput, phenotypic analysis as well as analytical tools.

But what we are bringing into this mix now is the semi-targeted solution.

So, these are basically transposase-based elements that these are mobile elements that allow for better identifying of better areas where transcription would be improved, essentially. And by using this technology, what we are seeing is, if you look at the rightmost plot where we are comparing with the excess of our random integration versus what we are getting in terms of titer with semi-targeted integration, significantly better tighter, also, less copy numbers.

And some additional data from the the leftmost plot is essentially showing random integration for the same set of proteins. And then the transposase platform when we are in small scale with our standard media. And when we are going to the CELLiST basal media, which is, a KPI developed media, produced by Ajinomoto. It's off-the-shelf media now.

When we move to that media, we get even higher titer, essentially. So, as is, we are getting improved productivity.

And by moving to the right media and feed platform, you know, we are getting eight to ten gram per liter, in many cases.

We are also showing the product quality between the random versus a transposase-based platform, and it's very consistent. There aren't any biases.

And, also importantly, we see high stability of these cell lines.

So, some data from scale up. So for one of our transposase cell lines, we went from three liter to two hundred liter scale. And the data we are showing here is the high consistency between three liter and two hundred liter. And the productivity achieved here is about 11.6 gram per liter in fourteen day culture. And the bottom panel is showing all the product quality attributes, essentially, again, showing consistency across the scales for the transposase platform.

On top of that, for the same cell line, when we are using our own feed media, we are currently developing a feed media.

As I mentioned, we have a basal media in the market already.

What we see is with that feed, the current version of the feed, what we got in two hundred liter with the commercial best case feed was eleven point six gram, and we are getting about eighteen gram per liter by using the feed we have developed inhouse. So, significant opportunity here for further improvement in productivity.

So, with the transposase line, not only are we getting better consistency, better productivity, but we are also moving faster. Actually, it's giving us faster timeline. So what we are showing here is with this platform and the staggered way of development, we can go from transfection to material generation for IND-enabling tox under two different scenarios. One scenario is if you are using the pools, pool of the top clones, which lot of customers are very comfortable doing. It's it's a risk-based decision.

Then if we are combining the top clones and making that into a pool within five months, we can have the tox material. And as we all know that tox is usually rate limiting, not so much the IND and GMP material generation, because the tox timeline is fairly significant. Right? So being able to go to tox fast with the platform that will give you robust and stable cell line is a big advantage.

And if we want to go with top clone instead of the pool of clones, then, it is about six months for us to go from transfection to the material generation.

So a little bit about our afucosylated cell line.

This cell line essentially was developed using our CHO-M line and subsequently doing the sequence of activities that's shown here in terms of sub-cloning and then, using the lectin assay-based glycan profiling to identify the right clone, right host cell, that will allow us to get afucosylated proteins.

And we used a particular licensed gene editing tool for this.

And what this data is showing is we had three different afucosylated host cell lines that we identified.

These are shown in these three different blue, green, and orange colors. And it's compared to our host cell standard host cell CHO-M line. And what we see is consistent growth, consistent glycan profile of two different types of proteins, except, of course, we get the afucosylated profile by going to this, fucosyl transferous knockout cell line.

Many of our customers have this need for their ADCC-based products. So we believe that this is a great offering, especially because of the high productivity we are able to achieve. A lot of the cell lines in the market for a fucoxolated offering don't achieve the right growth or productivity, so this is a big advantage.

And the last offering is this transient platform. What's nice about it is, again, it's based on our CHO-M, platform, so you are getting the identical background.

And, some of the product quality data at the bottom is showing, essentially, the stable expression versus the transient are extremely consistent.

So within a month you can generate material for either your candidates or for a particular molecule, one candidate that you want to quickly generate material on. And this is a great option where easily we can get a gram per liter kind of titer in a transient setting.

So with that, in conclusion, the new transposase line is giving us high productivity and highly stable cell lines with shorter time frame. And, the shorter time frame translates to transfection to IND-enabling tox in five months if we use pool off clones and six months if we use the final clone. And then a transient transfection, candidate selection and, PD start through it is a great advantage. I think it does meet a particular need in the market, and the afucosylated cell line is showing great promise as well.

And with that, I thank you all. Thanks.