0:01 My name is Christian, and I'm the managing director at NUMAFERM.
0:05 We are working on peptides, proteins, and something we call pepteins.
0:12 Pepteins are a term you might not be familiar with.
0:17 Let's start with an introduction to this term.
0:29 Maybe a different one here.
0:41 OK, you just wait.
0:48 Ah, here we are.
0:50 Let's start my presentation with the definition of pepteins.
0:56 Hopefully, that's acceptable for you.
0:58 Peptides are molecules containing not more than 50-40 amino acids based on FDA nomenclature, still considered small molecules.
1:13 Proteins, on the other hand, are typically long variants of peptides, consisting of 150 or 200 amino acids, making them fairly large.
1:28 Between peptides and proteins, there is a gap in the landscape, which we call pepteins. Pepteins have unique functionalities and are tough to produce because neither chemical synthesis nor recombinant technologies are easily adjusted for short proteins.
2:10 Within the peptein family, there are subfamilies like nano bodies, antibody fragments, disulfide-rich proteins and vaccines built off many epitopes. These are not easy to produce.
2:36 Our company has developed a new approach.
2:43 Our company is about the technology we have developed.
2:46 Let's jump onto the technology first.
2:50 We have developed a production platform called Numatech to produce peptides and proteins.
2:55 This platform allows us to produce peptides independently of the sequence, length, or functionality, and the processes we develop are very efficient.
3:10 The products we release are very pure.
3:13 Our production platform has two core technologies.
3:20 The first technology is called Numasec, which I developed during my PhD more than 10 years ago.
3:26 This technology allows us to fuse our target genetically to a Sectag protein and co-express a transport complex in the cell wall of Gram-negative bacteria like E. coli.
3:35 This process produces targets outside of E. coli.
3:51 We don't use this technology anymore at our company because it's a recombinant approach, which is slower compared to chemical synthesis. Being a recombinant process means its slow because you have to adjust the process to the target and the ideal environment of the target.
4:08 Despite its effectiveness, it couldn't meet the time expectations of our clients.
4:29 We learned that secreted Sectag proteins are highly soluble, while those kept inside the cells are not soluble and aggregate as inclusion bodies.
5:04 We found that these Sectag proteins contain GG repeats, calcium-binding domains, and the binding event converts the entire protein into a stable, soluble form.
5:54 We have shown that this system works for enzymes, very large enzymes up to 90-100 kilo daltons, being secreted functional. Outside of E.coli these proteins reach their final functional form.
6:11 Based on these observations, we developed a biochemical production platform. I will guide you through this technology step by step.
6:22 We fuse targets to a switchtag protein, produce inclusion bodies that are normally considered as waste, and refold the proteins in the presence of calcium ions.
7:11 Inclusion bodies have distinct advantages like high expression and purity, but the challenge is generating functional protein from aggregated proteins.
7:28 The primary structure of a protein is sufficient for proper folding if the environment is right.
8:13 We developed a technology to prevent intermolecular interactions during the refolding step. So we wanted to come up with a platform that grants access to the advantages of inclusion bodies while enabling the efficient, biochemical refolding of unfolded into functional proteins so that our technology would prevent intermolecular contacts during the refolding step.
9:04 We fuse targets to the switchtag protein, produce inclusion bodies, and refold them in the presence of calcium ions. The calcium ions then bind to the switchtag proteins, converting them quantitatively into soluble proteins.
9:25 The switchtag protein prevents intermolecular interactions, keeping the target in solution, allowing the target to fold properly.
9:53 This technology works for peptides, proteins, and pepteins (up to 600 amino acids).
10:14 Our clients didn't accept a switchtag protein attached to the target, so we developed a protease platform to cleave off the switchtag protein.
10:52 We based our developments on the TEV protease, which recognizes 7 amino acids and cleaves after the sixth amino acid.
11:56 Working with our partners we developed the Numacut TEV protease. They have developed enzyme that recognises the 6 amino acids but cleaves independently. This Numacut enables us to use our Numaswitch technology to produce targets independently of the sequence.
12:17 We released this enzyme as our one and only product, so this protease can be sourced on our homepage and through our distribution partners.
12:31 We produce inclusion bodies, refold them efficiently, and cleave off the targets tracelessly with the Numacut protease.
13:08 We train our cells to produce inclusion bodies containing the switchtag and the target, refold the proteins, and we introduce the Numacut protease to release the target from the switchtag protein.
13:30 The target is already very pure, making it easy to collect.
14:00 We have produced antimicrobial peptides which are not typically functional inside E coli but we switch them so that they become functional. The same holds true for growth factors, nano bodies, and fusion proteins with this technology. Even for highly complex targets this technology works because we give the target time to fold properly.
15:09 We handle short peptides up to 600 amino acids, with a standardized upstreaming and midstreaming approach. The downstream approach meets clients’ expectations in terms of scalability, we don’t typically rely on chromatography in this stage.
15:35 Production timelines are very short.
16:11 We have established protocols for state-of-the-art click chemistry and non-natural amino acid incorporation during fermentation.
16:45 We offer milligramme sampling within a month and gramme scale samples within a couple of months, with GMP material supplied within nine months.
17:13 We do everything in-house up to a 100 gramme non-GMP scale, with a partner network for larger scale production.
17:49 We work with pharma clients (approximately 60%), biotech, large pharma, and non-pharma clients from various sectors.
18:09 Our mission is to provide pure materials quickly with a scalable approach.
18:10 Thanks for listening.
