When Dyadic CEO Mark Emalfarb began the company, its focus was more centered on what people were putting on their bodies as opposed to in them.
“The journey can be described as going from ‘Jeans to Genes,’” Emalfarb told R&D Magazine/Pharmaceutical Processing.
The company took an intriguing path from supplying jeans manufacturers with pumice stones and enzymes to creating a proprietary protein expression platform based on a genetically modified strain of fungus that the company believes will replace Chinese Hamster Ovary (CHO). CHO is an epithelial cell line derived from the ovary of the Chinese hamster which is often used in biological and medical research and in the production of therapeutic proteins.
Making a Transition
In the 1980’s, Dyadic was at the forefront of the pumice stone industry. Clothing manufacturers would stonewash clothing with rough pumice stones to give them a used, faded look. Eventually, washing jeans with pumice stones fell out of practice and a new method took over: enzyme washing. So, Dyadic entered the enzyme sphere. As the company grew, Emalfarb started to wonder why his company was sourcing enzymes instead of making their own.
Emalfarb claims he had no business trying to develop and manufacture enzymes. “I was a journalism major in college. I didn’t know anything about science,” he said.
Fortunately, he was able to hire people who did. In the early 90’s, while developing enzymes for the purpose of washing blue jeans, Dyadic’s scientists discovered a filamentous fungal strain—Myceliophthora thermophile, nicknamed C1—which would ultimately hold more value than making jeans look worn-in.
“During development, two serendipitous mutations occurred,” Emalfarb explained. “The first changed the morphology of our organism, resulting in high productivity and better growth conditions. And the second created a strain that produced purer enzymes.”
It would soon be clear that C1 had potential to disrupt the bio-manufacturing industry. Over two decades, Dyadic built the knowledge, expertise, molecular tools, and technology needed to create and commercialize the C1 Technology Platform, a robust and versatile fungal expression system for gene discovery, development, expression, and production of enzymes and other proteins.
Advantages of C1
On average, the production cycle time of C1 is one-third to half the time of CHO’s, which translates directly to manufacturing and operational cost savings. Furthermore, compared to CHO, C1 can produce significantly more monoclonal antibodies (mAbs), which are the antibodies produced by a single clone of cells or cell line that consist of identical antibody molecules.
“Using an industry average taken from speaking with several of the top CDMOs, pharmaceutical, and biotech companies of 4 grams per liter in 14 days (336 hours) for CHO-produced mAbs—equivalent to a productivity of 0.3 g/l/day—Dyadic’s C1 technology already has demonstrated productivity levels of mAbs up to of 2.4 g/l/day, or nine grams per liter in 90 hours, or less than four days,” Emalfarb explains.
C1, which can be grown on low-cost, synthetically defined media, requires a smaller lab footprint than CHO to produce the same amount of mAbs, potentially transforming bio-manufacturing by allowing single-use fermenters—limited to 2,000 liters—to produce a similar capacity as is traditionally produced in a 15,000 liter fermenter.
Another incredible characteristic of C1 that Emalfarb said will save manufacturers time and money is its ability to secrete proteins naturally—an ability that CHO cells lack.
“C1 is a filamentous fungus that naturally secretes proteins, meaning you don’t have to force or initiate secretion by using expensive media,” Emalfarb said.
C1 does not only produce more proteins at a faster rate, but the proteins it does produce are also significantly purer than the ones typically produced by CHO.
“Dyadic has developed C1 cell lines that have eliminated a great portion of background proteins being produced naturally from the C1 cells,” said Emalfarb. “We have up to 80 percent purity of the secreted protein target. In the case of antibodies—where we believe we still have further room for improvement—we are already seeing levels of around 50 percent of the secreted protein being the protein target of interest, such as a full-length monoclonal antibody, as well as the certolizumab antibody fragment we are producing at 12 g/l in 112 hours with nearly 50 percent of the secreted protein being the antibody fragment itself.”
The combination of high yield and high purity, along with the ability to eliminate two steps in the purification process, due to no viruses being present from C1-expressed proteins, has the potential to make the downstream processing simpler and faster, as well as reduce manufacturing costs.
Emalfarb said that C1 has attracted a great deal of interest in the pharmaceutical industry. The company is involved in funded research collaborations with some of the largest pharmaceutical and biotech companies such as Sanofi-Aventis and Mitsubishi Tanabe Pharma.
Additionally, the company is currently working with both the European Union in the Zoonoses Anticipation and Preparedness Initiative program (ZAPI) and the Israeli Institute for Biologic Research (IIBR) and, as recently as this week, attended the Biomedical Advanced Research and Development Authority (BARDA) Industry Day conference.
A few months ago, FDA commissioner Scott Gottlieb’s stated, “Less than two percent of Americans use biologics, but they account for 40 percent of total spending on prescription drugs. They also represent 70 percent of the growth in drug spending from 2010 to 2015 and are expected to be the fastest-growing segment of drug spending.”
Emalfarb agrees that these numbers are troubling.
As an example of how the C1 gene expression platform may be able to disrupt the biologic industry, Emalfarb references the struggle to get biologic vaccines and drugs to patients is Hepatitis B (HBV) in China. As of 2006, China, where one-third of the 350 million individuals worldwide infected with HBV reside, has immunized 11.1 million children in its poorest provinces as part of several programs initiated by the Chinese government.
“However, the effects of these programs have yet to reach levels of immunization that would limit the spread of hepatitis B effectively,” he said. “We have to do better, and in order to deliver accessible and affordable healthcare to patients while reducing the financial burden on the healthcare system, we need to swap out CHO cells with cell lines that can speed up the development and lower the cost of biologics.”
He continued, “In order to truly disrupt bio-manufacturing, it seems that there needs to be a changing of the guard in the cell lines being used in vaccine and drug development and manufacturing.”