In a lab in Alberta, Canada, hundreds of thousands of fruit flies are stored in proprietary containers to produce kilograms of recombinant proteins for research and, as just announced, clinical use. Future Fields, a biomanufacturing company, is using the fruit fly (not fruit fly cell lines, but actual whole fruit flies) to produce recombinant proteins for in vivo CRISPR-based gene therapy developed by preclinical biotech company Jenthera Therapeutics in a sustainable and cost-efficient manner.

“Drosophila is literally the oldest laboratory model organism in the world, and we rear them at large scale using some novel techniques that give us some tremendous advantages around cost and scale, like fully continuous production,” Matt Anderson-Baron, PhD, CEO and co-founder of Future Fields, told GEN Edge. “Then we harvest the larvae and get the protein down to clinical-grade purity, same as you would with any other system.”

The two Canadian companies have announced a collaboration to use Future Fields’ EntoEngine, the world’s first synthetic biology system that uses fruit flies for recombinant protein production, to develop a first-of-its-kind cancer-fighting protein. Jenthera Therapeutics’ hybrid molecules are made up of cell-specific nano-antibody-binders (NABs) and CRISPR ribonucleoproteins (RNPs).

“For the stuff that we are trying to take through to the clinic, we are working with the microbial expression systems that will do a job, but you’ve got to seriously ask questions about the cost of goods sooner or later,” Philip Roche, PhD, CEO of Jenthera Therapeutics, told GEN Edge. “The objective is not to be a million dollars per treatment drug. And the best type of sustainability doesn’t just solve for energyit solves for cost and delivery to the patient.”

“We’ve got to drive down costs while maintaining quality and using innovation to achieve that,” said Roche. “Continually seeking innovative platforms that can produce at scale, at cost, and are energy efficient is absolutely essential, particularly when we consider the going rates of not only biologics globally but also gene therapy and gene editing systems as they get closer and closer to the market. This is really the potential of Future Fields’ EntoEngine.”

NABbing cells

The identification of a tumor-associated antigen enables cell-specific targeting for treating cancer. Jenthera has been working on an EGFR-targeted NAB to ablate oncogenes. The system can also be used to generate CAR-T cells in vivo.

Jenthera’s NAB technology behaves like a monoclonal antibody that binds to receptors on cells and shuttles in the CRISPR RNP. These fusion proteins are highly modified with loads of bells and whistles to deal with immunogenicity, stability, and endosomal escape.

Roche said that Jenthera is at the forefront of intravenous injections of CRISPR RNPs because they design their recombinant proteins to deliver in vivo without using lipid nanoparticles (LNPs) or viruses. Roche said that LNPs are not very stable and that while viruses can be very stable, they have their own targeting and manufacturing issues. “We’ve modified the proteins for the purposes of delivery and can tailor their in vivo half-life after intravenous injection,” said Roche. 

Jenthera’s lead, which was the basis for the collaboration with Future Fields, is a NAB-RNP for targeting EGFR-positive non-small cell lung cancer (NSCLC). “We ablate these cancers by targeting mutant KRAS and have demonstrated the in vivo shredding of tumors,” said Roche.

Once Roche and Jenthera got their hands on a great construct that showed proof of concept in animal models, they began to look at the most innovative ways to produce it and how to scale it up. Roche said that while microbes are great for making proteins during drug discovery, they are expensive and time-consuming. “Matt and his company have unparalleled scalability, but that’s also true of E. coli expression systems,” said Roche. “The leap that Matt’s giving us is the opportunity to do it at a better economic rate, better use of energy, and more rapid scale up.”

And that’s exactly what brought Roche and Anderson-Baron together. “If you’ve got a metric ton of biomass, even if the expression is moderate in scale, the EntoEngine is going to outperform biofermentation,” said Roche. “Wet biomass for wet biomass, the EntoEngine is always going to produce more.”

(Fruit fly) protein shake

Anderson-Baron co-founded Future Fields to produce a wide range of recombinant proteins that overcome the limitations of existing cell-based methods. “We genetically engineer whole insects, rear them at large scale, and then use them as a vehicle for the production of recombinant proteins that are used in a wide variety of applications,” Anderson-Baron told GEN Edge. “In this case, we’re talking about this novel protein that has some really exciting applications in the clinical setting.”

Anderson-Baron explained that the entire process begins with modifying fruit flies via microinjection into Drosophila embryos, a process that has been done for decades and can be outsourced. After receiving their genetically modified flies, the company can begin screening strains to determine which ones yield the best expression, after which they can rapidly scale up production. “We can scale to metric tons of biomass within a couple weeks,” said Anderson-Baron.

The manufacturing at Future Fields, which occurs in Edmonton, can continue as long as the flies are inbred to keep the genes in their progeny. “It’s cyclicalevery day we can harvest biomass and store it away, which is very different from cell-based systems where you’re doing batch-to-batch production, which can take weeks,” said Anderson-Baron. “So, we don’t have these large, weeks-long production batches. It’s just a continuous process. You’re maintaining that gene basically in perpetuity.”

Anderson-Baron, who is CEO at Future Fields, said that they haven’t discovered the boundary conditions of the EntoEngine. “We’re a three-and-a-half-year-old company, so we haven’t had the time to fully explore that and see what proteins we can and cannot produce,” said Anderson-Baron. “What I would say is that we have a lot of genetic tools to play with in our system. So in the event that there are challenges with a particular protein, there are tons of disposable tools to circumvent those challenges.”

According to Anderson-Baron, they already have use cases for doing just that. For example, he said that Jenthera’s NAB-RNPs were particularly challenging at the start, as the recombinant proteins were killing the insects. But Anderson-Baron said that they have a variety of ways to fine-tune the expression levels to allow the insects to be more viable.

“We have so many tools to address it that I think the variety of proteins we can produce in our system extends beyond any other platform and instills the advantages of peak cost-effectiveness.”

This partnership will help Future Fields expand beyond its initial set of products for sustainable, serum-free growth factors, such as human and recombinant bovine FGF2. With these two products, Future Fields recently achieved the esteemed ACT Environmental Impact Factor Label from My Green Lab, a nonprofit organization with the mission to build a global culture of sustainability in science. As the first growth factor in the world to obtain the premier eco-label, the ACT Label highlights Future Fields’ dedication to producing sustainable recombinant proteins.

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