From The Editor | June 26, 2015

DARPA's Challenge: Manufacture A Biopharmaceutical In Less Than 24 Hours

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By Trisha Gladd, Editor, Life Science Connect

Biologically-derived Medicines on Demand (Bio-MOD)

American soldiers fighting in remote places overseas do not always have the luxury of picking up the biopharmaceutical they need at a nearby pharmacy or hospital. On the Defense Advanced Research Projects Agency (DARPA) Battlefield Medicine site, Dr. Geoffrey Ling, a retired Army colonel and program manager at DARPA, writes that it can take weeks to months for these drugs to make it to battlefield frontlines. Oftentimes, this can be too late for the person who needs them.

To address this critical gap, DARPA created two programs for developing devices and techniques to rapidly produce pharmaceuticals on demand. This includes in warzone situations as well as disaster situations, such as the insulin crisis that occurred in New Orleans following Hurricane Katrina. One of the initiatives, Pharmacy on Demand (PoD), is focused on small molecule drugs. The other, Biologically-derived Medicines on Demand (Bio-MOD), is for protein-based therapeutics. “Both PoD and Bio-MOD efforts will seek to develop platforms for manufacturing single-dose levels of FDA-approved APIs and biologics and demonstrate high purity, efficacy, and potency in short time frames,” explains Dr. Ling on the site.

There are currently only three research teams whose proposals for the Bio-MOD project were awarded funding by DARPA, each with a different idea on how to approach it. I spoke with some members of one of these teams, made up of experts from the University of Maryland, Baltimore County (UMBC), Ohio State University (OSU), Thermo Fisher Scientific, and Latham Biopharm. The team’s founding member and principal investigator is Dr. Govind Rao, professor of chemical, biochemical, and environmental engineering at UMBC and director of the university’s Center for Advanced Sensor Technology. In 2012, he attended a public briefing where Dr. Ling described the challenges of getting medicine to the field in remote locations. This briefing eventually led to the creation of the Bio-MOD program.

A Biopharmaceutical Company In A Laptop

Dr. Rao confesses that when he went to the initial DARPA meeting for the program, he was skeptical something like this could ever work. “It takes 24 hours to even grow cells, let alone produce anything from them in less than 24 hours,” he explains. However, he says he was leafing through a trade magazine shortly after the meeting and saw an ad from Thermo Scientific. It talked about the company’s mammalian cell free-expression system and its ability to produce even large proteins within hours. This cell-free extract is also available in a lyophilized formulation, which eliminates the need for cold chain shipping and could advance the project even further down the road. He refers to this as his “eureka moment.” Dr. Rao contacted Thermo Scientific, and they eventually partnered for the Bio-MOD proposal. This was the first step in building what later became a team of seven members.

Because the goal of the DARPA project is to make and purify proteins in a device the size of a laptop, Dr. Rao needed somebody who could address the need for protein purification. Through academic biotechnology journals, he was already aware of the innovative work by Dr. David Wood, associate professor of chemical and biomolecular engineering at Ohio State University. Dr. Wood is known for his research in the purification of recombinant proteins using a self-cleaving affinity tag technology based on small proteins known as inteins. As he describes it, inteins are mobile genetic elements that were discovered shortly before he started grad school in the mid-90s. Inteins (intervening proteins) are polypeptides that are found embedded in the middle of other proteins in nature. Once translated into a mature protein, inteins exhibit the remarkable ability to excise themselves from their host proteins through a self-splicing process.

By re-engineering the intein’s self-splicing reaction into a self-cleaving reaction, Dr. Wood and his advisors generated a new tool for protein purification. Unfortunately, because of a broad competing patent on another intein design, he was prevented from commercializing his design at that time (it is still not commercially available). However, he has shown in several peer-reviewed papers that his self-cleaving intein is highly useful for purifying many proteins with a variety of self-cleaving tags. It is currently in use by dozens of other academic laboratories all over the world, which is why Dr. Rao contacted him for the DARPA project. “Because my technology is applicable to practically any protein, it would be a really powerful platform for making this [Bio-MOD device] work,” says Dr. Wood. “I reviewed Dr. Rao’s proposal and determined that we could definitely use an intein for the Bio-MOD project and effectively have a self-cleaving affinity tag be the first capture and concentration step for purifying all of the different proteins we would want to make.”

Dr. Rao’s Bio-MOD team is looking at an in-vitro translation system with the ability to produce complex glycosylated proteins. They will then use Dr. Wood’s intein-based purification platform to purify whatever protein is produced in the device. From there, some polishing steps will be developed, and a QbD approach and advanced process analytical technology (PAT) will be used to assess and ensure quality. Dr. Rao says the project includes a system design team to focus on the hardware, a protein expression team optimizing the expression system, a purification team to figure out how to purify the protein, and a regulatory team that offers guidance from industry advisors. The regulatory team includes partners from Merck, Pfizer, and Johnson & Johnson.

A Long Road Ahead

The Bio-MOD project was divided into two phases. The first phase, which was recently completed, required a proof-of-concept to show that the team’s basic ideas would work. The second phase involves making the first integrated device that can produce six different model therapeutic proteins in a fully-formulated and ready-to-inject form. This is a challenge because in conventional large-scale systems, there is plenty of workaround and troubleshooting without any issues of water supply or pressure to push materials through a column. All of that changes when you are working at a millifluidic scale. An unexpected air bubble could choke the whole flow and stop the system, while being practically invisible to the operator. 

“We are entering a new realm of figuring things out,” says Dr. Rao. “Everything that happens today to manufacture a biopharmaceutical has to be miniaturized into this briefcase-sized device. There will be little chambers that are potentially at different temperatures with small amounts of fluid pumping through, all while ensuring that you don't end up with something like running out of buffer. We are working through all of the calculations to understand any potential issue, in order to make sure that the scaled-down process is representative of what you successfully did in the lab and operates in a reproducible fashion.” He adds that this technology would also require regulatory authority approval, and the team has started discussions with the FDA to ensure that regulators are engaged early in the development process.

A Disruptive Innovation

While the success of this project would certainly improve our ability to treat soldiers in the field or victims of natural disaster, it also would have a major effect on today’s pharma business model. The entire industry is looking for the most efficient and safest way to manufacture drugs. If Dr. Rao and his team can successfully build a system that has the ability to create an on-demand biopharmaceutical in less than 24 hours, would the large facilities of today even be necessary? The answer to that question is still very far away, but it’s definitely one that comes to the surface as the DARPA teams continue to move down this path.

Today, biopharmaceutical companies and regulators are also exploring different ways to bring drugs to market that do not have large commercial interest (i.e., orphan drugs). In this realm, the Bio-MOD system could thereby serve as a game changer for applications in personalized medicine. The beauty is that the economics become the same for individual doses for each product, whether it’s a blockbuster drug or a rare protein only a couple of hundred people need. What does the industry look like when market size becomes a non-issue?

Another area where Dr. Wood thinks Bio-MOD could have an impact is research. With this system, a scientist who discovers a new gene/target or wants to create a new drug can test it quickly. “Instead of having to create a completely new cell line for each protein, they can go straight from gene to purification of their target, and then find out if it’s active, amplify it, tweak it, and test it again,” he explains. “This device would allow them, within 24 hours, to have enough of a protein that they can experiment with it.” The ability of academia to quickly determine the potential of a therapeutic target that is still in the very early stages of testing would be huge for an industry that often looks to them for innovation. High-risk work such as this is often too much of a financial liability for many pharmaceutical companies. However, with enough funding, a university scientist can discover breakthroughs that today’s industry might never fathom.