1. DARPA awards CAST $8M for new phase of
drug delivery project
June 24, 2015 1:44 PM
By Sarah Hansen ’15, M.S., Biological Sciences
The Defense Advanced Research Projects Agency (DARPA) recently renewed a grant for
approximately eight million dollars over two years for a team at UMBC’s Center for Advanced
Sensor Technology (CAST). The team is developing a portable device the size of a briefcase that
can produce therapeutic proteins, such as insulin, in only a few hours and in small batches. The
device would be critical in situations where medical supply lines have been cut, such as in war
zones or following natural disasters. The project is known as BioMOD, for “biologically-derived
medicines on demand.”
The new device, which could eventually replace the current centralized model of pharmaceutical
production, is like “going from a mainframe computer to a laptop,” said Govind Rao, CAST
director and professor of biochemical, chemical, and environmental engineering (CBEE) at
UMBC. “It empowers people in ways that are unimaginable,” he said. The device would first be
used in hospitals, but the team’s vision includes eventual home use.

2. UMBC is the lead of a consortium including the Ohio State University (protein purification),
Thermo Scientific (cell-free expression system), and Latham BioPharm (system integration), and
there are approximately 30 team members across all sites. UMBC teammates come from several
departments: CBEE, Computer Science and Electrical Engineering, Mechanical Engineering,
Chemistry, and Biological Sciences.
During the first two years of the grant, the team showed that their general approach can work. A
bioreactor approximately the size of half of a soda can contains cellular extracts from Chinese
hamster ovarian (CHO) cells. CHO cells are “an industry workhorse for producing
pharmaceuticals,” said Rao. The beauty of this device is that the cellular extracts are “essentially
the cell minus the nucleus,” Rao said. That means the tricky task of keeping cells alive isn’t
necessary, but the protein-production machinery is ready to go when one adds DNA coding for
the desired protein product. The bioreactor builds the desired protein in two to four hours. The
short time scale also reduces contamination risk. The next step is purification, where you “fish
the product out” of the cellular slurry in the bioreactor, Rao explained. The third step, polishing,
is an even finer process to remove any remaining impurities.
The team knows the device works, so “now it’s the real deal to show that it works in a robust
enough fashion to produce molecules of the necessary purity to safely administer to a human,”
Rao said. The team will be carrying out “an exhaustive amount of validation” to show that the
protein product is of similar “purity, potency, and structure to that made by a conventional
commercial process,” Rao explained.
This next two-year phase will stop short of clinical trials, however. The team is looking for
commercial partners to help support extraordinarily expensive clinical trials once they
thoroughly validate the prototype and procedures in the lab.
Rao appreciates working with DARPA. “They are not afraid of risks, and they have an
extraordinary tolerance for failure. That allows us to try bold things that we ordinarily wouldn’t,”
he said.
That tolerance for failure has certainly been tested. This spring, a key piece of equipment used
for validation had consistent problems. It required a major overhaul and new training for CAST
team members, plus it was out of operation for several weeks. I experienced the frustrations of
the scientific process firsthand in my role as a CAST graduate assistant whose primary
responsibility was this particular instrument. Before I left, though, the machine was up and
running again. It hadn’t produced any important results yet, but Rao was quick to emphasize that
“every little bit is important. It’s all about the team.”
It has its challenges, like all worthwhile endeavors, but, “This is going to be making history,”
Rao said. “Someday 20 years from now, when you’re injecting yourself with a drug you made
yourself, you’ll say, ‘I was there.’”
Source:http://research.umbc.edu/umbc-research-news/?id=52413

4. DARPA's Challenge: ManufactureA
Biopharmaceutical In Less Than 24 Hours
By Trisha Gladd, Chief Editor, BioProcess Online
Follow Me On Twitter @bioprocessol
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

5. 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 OSU. 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

6. 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

7. 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.
Source:http://www.bioprocessonline.com/doc/darpa-s-challenge-manufacture-a-biopharmaceutical-in-
less-than-hours-0001