November 17, 2023

Top challenges in recombinant protein purification process development

In this video, Emma Lind, Product Manager for Chromatography Resins at Cytiva answers questions from BioPharm International about the challenges faced by process developers working with recombinant proteins.

Cytiva™ Protein Select™ resin is designed overcome some of these challenges.
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Jeanne Linke Northrop: Hello, I'm Jeanne Linke Northrop with BioPharm International, and I'm joined today by Emma Lind, Global Product Manager for resins and technologies with Cytiva. Emma, welcome.

Emma Lind: Thank you. Thank you for having me.

Jeanne Linke Northrop: Oh, absolutely. Thank you so much for providing me the opportunity to sit down with you. I'm really looking forward to this discussion today on recombinant protein purification process development.

In the industry, we're seeing recombinant proteins more often, right? And we're seeing some common challenges of course arise when it comes to exploring these more innovative and potentially lifesaving modalities that are starting to change the world of medicine. I know they're being utilized and studied for various diseases. So I'm really looking forward to talking more about some methods that can be used to help navigate this complex field, as well as address some of these concerns that are being raised by scientists. I'd like to pass the floor over to you, Emma, and ask for some insight here. First of all, why can process development be much more complex for recombinant proteins compared to traditional monoclonal antibodies?

Emma Lind: Yes. Well, for recombinant proteins, you often don't have an affinity solution, an affinity chromatography solution that can be used, which is very different from mAbs, which do have a great affinity solution that can be used in protein A, such as Cytiva MabSelect PrismA. But then there's also a good platform approach that can be used. So after your affinity step, you can use a combination of ion exchangers and the monoclonal antibodies will react quite similarly each time, so this platform approach can be used. Whereas for recombinant proteins, we see a great uptake in the different kinds of recombinant proteins that are coming out. And each of these proteins have a different property, even a protein, a recombinant protein that's very similar to each other will behave differently and need a different solution. And then there's the wide breadth of different kind of recombinant proteins that are being purified from enzymes and peptides. Quite a lot of different kinds of scaffold proteins and other kinds of recombinant proteins that are going to need different solutions. So you can't use this platform approach when purifying them.

Jeanne Linke Northrop: Thank you, Emma. And talking about not only the need for different solutions, but it just really, really emphasizes just how unique recombinant proteins are in that they all differ. It's very challenging. So in addition to that, as I would feel would be a major challenge in process development, maybe you can highlight some other challenges that are being reported by scientists?

Emma Lind: Absolutely. So these recombinant proteins, we see several different challenges that process developers are facing. The number one challenge they face is that they're unable to get good purity in their first step. If they could use an affinity solution, they could get more than 95% purity already in the first step. But these recombinant proteins do not have an affinity chromatography solution that would work across all recombinant proteins. So they have to use a combination of different techniques, and especially in this first step, they're not going to get the purity of this level.

Another challenge that they face is this slow development time that can take months even, where since they need to use a different combination of methodologies such as HIC and ion exchange, each of these different processes take different optimization that needs to be done for each process. And we see even ourselves that the different combination of processes can cause different considerations that need to be taken. Sometimes you need to, if you find a good solution in the first step, you need to change your procedure in the next step to make it work. So there are a lot of different aspects in the process development time that need to be taken into consideration. And we see that process developers, when they see a molecule that just looks too complicated to take in, they're going to shelf it and not even consider it in the process development.

And then the other thing that is a challenge for process developers is that the actual process itself of purification can be quite lengthy. As I mentioned, they need to do a combination of different steps such as ion exchange and HIC, but they might also have to add non-chromatography methods, such as acid precipitation, filtration, other kinds of things that need to be taken into consideration and take time in the process itself. And, of course, this time in the actual purification of the molecule adds to added cost. So, these can be quite a big challenge for process developers when they're setting up a procedure for the purification.

Jeanne Linke Northrop: Thank you. And despite all of these challenges though, we do know that these are potentially safe, and they have a lot of potential in the industry. So that leads me to my next question, Emma. How does Cytiva help process developers address these current challenges that you mentioned? Because again, speed, cost, are major demands in the industry right now in drug development.

Emma Lind: Well, we at Cytiva are looking at different technologies that can help to provide an affinity chromatography solution for molecules that don't have a solution today. One example of this is a technology that I've recently presented at PepTalk and PEGS, that is actually a tag affinity solution that will allow for customers to use this product but not have any amino acids left after purification, which is quite different from any tag on the market today. And a reason that process developers and drug producers do not use this today is because the traditional tags leave amino acids on the protein and can use some chemicals in the cleavage of the tag from the molecule that are not conductive to large-scale purification. But this is a solution that we're looking at to provide, where it solves these hurdles by having a tag that can be used in a way that does not leave any amino acids left on the protein, and also does not use any special reagents for the cleavage of this.

Jeanne Linke Northrop: Thank you. Very interesting. I briefly saw some information on the website about that, and I also found a lot of helpful resources, specifically on recombinant protein study as well. Emma, again, thank you so much for your insight today. I appreciate you taking some time out of your very busy day to meet with me, and I really appreciate speaking with you.

Emma Lind: Thank you for having me.

Tags: process development, Recombinant proteins