0:00
The first of whom is Fabien Rousset, who is the Director of Global innovation of PolyPeptide.
0:08
He has a very impressive CV in the industry, 25 years of experience in the biopharma industry with a PhD in polymer chemistry.
0:19
And he has applied this particular skill to low pressure and HPLC consumable equipment from batch 2 continuous.
0:30
He had developed chromatography and filtration techniques for small and large molecule with the main set of process intensification.
0:40
So Fabien, thank you and we'll look forward to your talk, the title of which is our solution for greener and controlled processing.
1:07
I'm not a singer, so I don't know how to use the microphone, but OK, I can try.
1:13
So as you were mentioning, I'm coming from the biopharma or the mAb industry.
1:17
So that's clear that coming from that field with low pressure with single use technologies and arriving at PolyPeptide as a global director is a major change.
1:27
So please be kind if I'm saying some mistakes.
1:31
I'm there since only two months.
1:33
So as we are an innovative company, they are innovating by putting me on the stage after two months or so.
1:37
I will introduce the polypeptide group and a bit of messages regarding the green programme that has been induced in the last years, some projects internally to address those green consideration.
1:56
And I will go through four major solutions that we have.
2:00
We have worked on especially addressing the PFAS, green solvents, a new concept for improving the use of some resins and also the use of advanced control software to be sure that we are controlling the process very well.
2:16
And the conclusion.
2:17
I will have also some games within the presentation just to be sure that you are following.
2:23
So listen carefully, please.
2:25
So, OK, you probably know PolyPeptide more than myself.
2:29
So the company's existing since more than 70 years originally from Sweden.
2:34
So they have expanded by several acquisitions now covering all three major regions like US, Europe and Asia.
2:42
And the last acquisition or a major point was in 2021, the creation of headquarters in Switzerland. Covering the planet means we have several sites with some specificity particularly in California in Torrance, in close to LA and in San Diego.
3:01
We also have in Europe three major production sites in Malmo, the first one in Braine-l’Alleud in Strasbourg as well in France where I'm located.
3:13
And we also have one in India specifically associated to generics.
3:20
As you can see we are 12 more than 1200 employees worldwide and we have a substantial number of projects per year which is more than 200 and all the sites are GMP certified. In Baar, this is the HQ.
3:33
So these guys are not making any production.
3:38
Our core values, so as I was mentioning is as a preamble innovation, this is clearly something that is really included in the mindset at PolyPeptide.
3:47
We really want to innovate, across cross functionalizing the expertise within the company always willing to raise the standard of production.
4:00
So by introducing new innovation solutions, the excellence and trust is always, a key message that we want to provide in all projects that we have with customers, but also through some collaborations.
4:12
We're having a lot of collaborations with academia and partners and companies.
4:18
This brings me to the classic business model.
4:20
And we are CMO for peptide.
4:22
So starting from small scales to large scale GMP, non GMP to GMP.
4:27
And we really accompany all our customers, all our collaborators through the, the early phase to the late phase, supporting them by production, but also on the analytical side of it.
4:39
And from a regulatory standpoint also this last point is very important nowadays because we do have more and more pressure to produce on the greener side, of the process.
4:55
What does it means?
4:56
For sure you have already seen this environmental social KPI that's clear that at PolyPeptide we are more focusing on the chemistry with the green chemistry.
5:07
Also the responsibility to be sure that those new technologies are applicable from small scale to large scale and also in the supply chain engagement.
5:16
And this is derived into some key points that we are having at PolyPeptide, which is generally you used to see those reduce, replace, recycle or avoid chemicals.
5:28
This is really the key things that we are having in all projects that we are running right now, and we will go through the project keeping this in mind.
5:41
The first one I would like to discuss is the PFAS free.
5:45
So that's clear that this is a new regulation here.
5:48
It came by the angle of cosmetics.
5:51
So at PolyPeptide this is also very important topic.
5:54
So we worked a lot on the on this PFAS-free, especially since the TFA which is dominantly used for the cleavage is used and considered as a PFAS (per/polyfluorinated substance).
6:07
This is a very broad topic here and a very important one because of this dominance of the use, but also because it's spread in transversely in the processes and also in the different fields.
6:22
Because we also have seen recently that this product which is in phase 3, an anti-HIV product, has been questioned just because of this perfluoro groups on the product.
6:34
So it means that probably as a molecule in under development, we will not be challenged too much in the 2, 3, 4, 5 years.
6:43
But on longer term it's also important that we as producers and you as developers to take into account this PFAS new regulations.
6:53
Knowing that we have done a bit of bibliography for sure.
6:56
And if we consider the Bronsted acids and the Lewis acids it was well known as a single solution to have some properties for cleavage.
7:07
And so we thought that probably we could develop a new approach based on that for replacing TFA.
7:13
This is what we have done here Fmoc-Leu on some resins, HMPA AMS resins, polystyrene resins.
7:20
So what we have done here is we have tried to use iron chloride in some different solvents.
7:27
And finally at room temperature, we could see that the yields were not really good.
7:31
So we had the opportunity and the idea to combine different Bronsted acids and Lewis acids.
7:40
And in that case, if you look here at 40°, we have quite good yields by combining iron chloride and acetic acid.
7:50
We do have the same, results combining also with chloric acid.
7:54
So this is clearly an indication that by combining the theoretical approach and a bit more innovative approach by combining different solutions strategies, we could develop an interesting TFA free solution for cleavage.
8:09
So question who is represented on that picture, Laa-Laa, who is a Teletubbies for the people who doesn't know at Peptide lab, me at Rio’s carnival operator when handling TFA or Elon's dress for Mars.
8:21
So everybody knows that this is operator when handling TFA.
8:26
So this is not ideal.
8:27
And for safety reasons, the regulation is also trying to reduce the use of TFA.
8:33
The second topic, which is still in correlation of the first one or can be in correlation of the first one.
8:39
We worked on the replacement of DMF.
8:41
Sorry, many presentations by Lorenzo by you, Fernando this morning about the solutions to replace DMF.
8:50
That's clear that we worked the last years on the on solutions to replace the DMF.
8:57
But the question is, do we just have to find a solution, a new solvent to replace DMF or do we have to have an holistic approach within the time and the more integrated, the more elaborate approach still in the mindset of having a great process.
9:14
Meaning if we use a green solvent but twice amount of green solvent or same amount of green solvent, it will not solve anything.
9:23
We have to get an integrated by adequate chemistry, minimising the consumption and still improving the process.
9:29
Even if we use a green solvent, we don't, we are not looking one step forward, but two steps forward.
9:35
So here what we did.
9:40
So we have here a first step of working with a green solvent.
9:45
And for sure it's not a very new regarding all the presentation that has been done before ethyl acetate and DMSO, we could do this ACP product, and we also have combined with a cleavage using the new solutions and we worked on different ratios of Lewis acid and Bronsted acids.
10:05
And for example, this combination led us to find good combination of green chemistry for the upstream and also a green chemistry for the cleavage.
10:15
And here we are defining some good recipe of solutions for peptides, which are tricky, to be made for sure.
10:25
We worked on green solvent.
10:26
One of the best green solvent is water if we do not consider the waste because the waste has to be also treated.
10:32
So we worked on the on an example of aqueous SPPS.
10:36
And here the idea was to combine with a cosolvent, and we have tried different cosolvent.
10:43
This is a long term project here because we are not there yet with the aqueous SPPS as you can imagine.
10:49
But it was also to see to which extent we can test the different green solvents.
10:55
It appeared that we can find here some rather good solution.
10:59
And for example, we are currently looking at the acetonitrile.
11:03
Probably this is not the best cosolvent in terms of chemical safety and chemical characteristics, but at least we are pursuing some tests right now on the combination of water and the acetonitrile for USP and DSP.
11:18
Because as you know during the downstream process and the purification particularly acetonitrile is also one reagent.
11:24
And when I was talking about the holistic approach when we developed a process, this is also a good point to reduce the number of reagents that we use in the process.
11:31
So if we can use during SPPS and during downstream the same chemicals, this is also from supply chains, CO2 footprints, an advantage.
11:43
So the next one I got already a question this morning about SPPS new resin, the Kraken concept.
11:49
So I will we'll have some slides for you here.
11:53
So what is Kraken?
11:54
It has been described since many years.
11:57
But for those who don't know here you have a classic SPPS resin.
12:01
So what we do instead of having one single point attachment where we put the linker and when the where the peptide is growing, we have a, we have developed a branching concept having a multi-point attachment of linkers in order to intensify or to increase the density of the growing peptide on the surface of the resin.
12:23
The resin is still the same, but prior to put the linker in to grow, we put a, we put, dendritic space, dendritic spacer to have a to have an optimised process.
12:35
If we look theoretical quantity of peptides and group peptides obtain this following, the trend.
12:43
We still have some slight variations.
12:46
But this is really interesting because we are not reinventing the wheel here.
12:52
We just take the basic resins, and we just put a spacer which is improving the growing peptide, the quantities, densities at the surface.
13:02
From a quality standpoint, we have good results depending on the different generations.
13:07
So sorry I did not explain.
13:09
The 1X, 2X, 4X is here you have the native resin.
13:13
Here we have a spacer which is doubling the growing sites.
13:18
Another generation on these two X which is doubling again, which means 4 active sites doubling again, 8.
13:25
And you can, if you want, you can do 8X, 16X, 32X.
13:32
How far can we go? Not to get with a problem of the steric hindrance or impurities, with deletions and so on. But the principles are exactly the same.
13:43
What about the resin?
13:44
What about the generation we have?
13:48
So here the question is we have to select the resin because if you still use one resin, how we can adapt this Kraken concept.
13:58
So select the resin nature, the resin, the generation of the Kraken, the reactor volume, because the resin will still swell.
14:10
You have also to understand that once we compare the native resin and the grafted one, so the Kraken derivative, we generally have a swelling which is much less than the native resin.
14:23
So if we consider also from a green aspect.
14:26
So we have an intensification of the process, which is good because we produce more with less, but we also have the same recipe in terms of washing and so on.
14:35
So one CV now is for example washing four times more peptide than with a native resin.
14:41
So this goes in the right direction.
14:48
Here you have a, you have an example of here this is a constant volume model. So 1X native resin 2X, 4X.
14:55
We keep the same amount of resin.
14:57
And here you have the quantity of peptide that can be produced with these resins.
15:01
You can see directly that the quantity is pretty much linear.
15:04
We still have now a plateau because of the steric hindrance, and we did not get the 8X for example would be much lower and so on.
15:11
But this is just to give you a visualisation of how efficient is this Kraken concept.
15:19
So branch.
15:20
So with a crude peptide we can see, sorry, with a traditional resin, we can see the Kraken one.
15:26
We have a very good, similarity between what we can produce with a native resin and what we can produce with a Kraken concept.
15:35
From a quality standpoint, there's no real bottleneck.
15:41
So which limitation do we have with a Kraken concept? For A, people raise your hand if you think that the peptide length is a problem with a Kraken concept? If it is a linker? If it is a resin? If none?
16:00
Oh I missed it, sorry.
16:03
OK now so far we didn't get too many limitations as I was explaining.
16:09
Steric hindrance is for sure occurring when all the vectors are growing.
16:13
So we are adjusting the generation of cracking concept.
16:16
Instead of having 8X and steric hindrance, we will probably just choose the 4X without steric hindrance.
16:23
So this is also a flexible parameter.
16:25
We have to always choose the optimal conditions and control the process behind that.
16:30
This is why we also have here a good thing.
16:39
The last one is the advanced control software.
16:48
So here the idea was, I was explained that traditionally we do with SPPS, we do the washing in batch conditions.
16:57
And that's clear that having a new way of doing the washings and probably also following the different kinetics of reactions.
17:05
We could potentially implement percolation protocol assisted by FT-NIR which is on the recirculation loop here.
17:16
And then we can follow the different steps of a process.
17:19
So coupling deprotection, washing, whatever, with this NIR in line.
17:27
And then by having a proper calibration in terms of when do we have to stop the step in terms of piperidine limit concentration and so on to start the next step.
17:38
Then the software here can tell you when to start the next step.
17:45
So if you have for example, sometimes a recipe which is based on volumes, you have to pass let's say 100 litres of washing buffer.
17:53
But if you know that after 80 litres it's OK, well you save 20 litres and if you repeat that among the, the different batches and so on, you have a greener process.
18:01
So this is really to optimise the washing sequence by improving the automation.
18:09
So here we have many parameters to play with and to control.
18:12
When once we define how the percolation should work, but it's also based on the what are the remaining species and what the limiting species as impurities when we start the next step.
18:25
So all these are taken into consideration, plotted, and then we generate the recipe and it can save to a substantial amount of buffers, more particularly for washing, which are generally low, poor efficiency performance.
18:40
So I was giving you some hints here with advanced control software, what is the maximum washing solvent reduction we can expect?
18:47
Raise your hand if you think that this is 10%? 10 to 30%? 30 to 60? or more than 90?
18:56
But sorry guys, this is not Christmas time.
18:59
So that's clear that this is 30 to 60% depending on the process, depending on the peptide, depending on the resin also you have.
19:06
So this is substantial, an improvement in terms of process.
19:11
So as a summary, if you don't want to be dressed like a Teletubby and to avoid using TFA, we can replace it by iron chloride acid mixtures.
19:23
If you want to participate and contribute to PMI reduction, that didn't give you too much numbers regarding PMI reduction, but we can discuss later on.
19:32
And we can also use the percolation to reduce the solvent consumption during washings. Having an intensified process that boost your productivity by 2X or 3X in terms of peptide per litre of resin, you can use the Kraken concept.
19:47
And if you care, if you want to have a better control and automatization of your process, you can also use the NIR in line.
19:54
But this is clear that all these solutions are provided by PolyPeptide.
20:00
And if you want to discuss more about that, please go to PolyPeptide booth #18 and regarding project and future collaboration, we are open to discuss.
20:08
I would like to thank at the end all these people, so from US, from Sweden, from France, only one from Belgium.
20:15
And he was drinking too much beer.
20:18
And I thank you.
