Epidemics and conflicts have significant implications for both raw material supply chains and the distribution of essential medicines. Doctoral candidate, Shayan Borhani, is part of a team at the Center for Advanced Sensor Technology (CAST) at the University of Maryland Baltimore County (UMBC) seeking innovative solutions to tackle these problems – might cell-free protein expression hold the answer?
Driven by the shortages of medicines during conflicts in Iraq and Afghanistan, then DARPA program manager Dr. Geoffrey Ling saw an opportunity to leverage recent advancements in protein production to bring the manufacturing of essential pharmaceuticals closer to those most in need. His vision materialized into the biologically-derived medicines on-demand (Bio-MOD) platform.
“In response to Dr. Ling’s vision, my thesis advisor Dr. Govind Rao came up with the idea of using cell-free expression systems in an automated fashion,” explains Borhani. “The idea is that you can express your product and purify it in a portable device and effectively bring those medicines to the battlefield.” The prototype of Bio-MOD is housed in a robust black plastic suitcase equipped with everything necessary to produce biotherapeutics. The entire process, from protein production to purification and finishing, occurs within a contained environment, thanks to the remarkable fluidics engineering behind Bio-MOD (https://doi.org/10.1038/s41551-018-0259-1).
“It’s similar to how you’d prepare your coffee in the morning, where you just add your pod, push a button and get your coffee,” Borhani continues, illustrating the user-friendly nature of the system. The enabling technology behind Bio-MOD is cell-free protein synthesis (CFPS). Borhani emphasizes, “The beauty of cell-free expression systems is their ability to generate your product in a very rapid manner. You are able to screen a variety of different candidate proteins more effectively than typical microbial or cell cultures.”
»You have to consider all the things you’re going to be supplementing into the reaction. The beauty of the ALiCE® system is that all of those components are built into the lysate.«
Shayan Borhani
Doctoral Student, UMBC
Borhani’s research focuses on the production of an antiviral drug, Griffithsin (https://doi.org/10.1016/j.nbt.2023.04.003), and the well-known diabetes management medication, Insulin. He highlights the motivation behind his work, stating, »There are a lot of diabetics around the world who ration their medication because they either don’t have access to a reliable supply or can’t afford it. We have the opportunity to develop something that can serve those patients and address this issue.«
Throughout his project, Borhani has explored various cell-free systems for the production of his candidate proteins, including CHO, E. coli, and the plant-cell based ALiCE® system. »When you’re looking at a cell-free system, you have to consider all of the things that you need to supplement into the reaction to generate your target,« Borhani explains. In the case of CHO, this meant adding a variety of different chaperone proteins to achieve enhanced expression of the therapeutic targets. For E. coli, strain engineering was required, combined with addition of chaperones – specifically for the formation of complex post-translational modifications (PTMs). »And then we came across ALiCE®,« says Borhani.
»One of the key reasons I selected the ALiCE® system is its ability to toggle between cytosolic and microsomal expression pathways depending on your needs for PTMs. And then you have to consider all the things you’re going to be supplementing into the reaction. The beauty of the ALiCE® system is that all of those components are built into the lysate.«
»Having the ability to just add your DNA and get your protein – and then in a matter of days, generating a purified product really is a distinct advantage. And I always got protein expression with ALiCE®.«
Shayan Borhani
Doctoral Student, UMBC
The simplicity and reliability of the ALiCE® system further contribute to its appeal. Borhani highlights, »Having the ability to just add your DNA and get your protein – and then in a matter of days, generating a purified product really is a distinct advantage. And I always got some protein expression with ALiCE®.«
Regarding the future of the Bio-MOD system, Borhani acknowledges that further testing is necessary before its deployment in real-world scenarios. Nevertheless, he remains optimistic: »Anywhere in the next 5-10 years, we’re going to start seeing this at home medical device where you’re generating your therapy on-demand. Once we finalize this technology, then we will really be fulfilling our goal of bringing therapies closer to the patient.«
ALiCE® is a eukaryotic cell-free protein expression system capable of producing complex proteins in under 48 hours. The proprietary cell-free lysate contains all of the machinery necessary to implement eukaryotic post-translational modifications, without specific optimizations and in a on-pot reaction.
In a world challenged by supply chain disruptions and limited access to essential medicines, the exploration of cell-free protein expression systems offers hope. Borhani and his team’s pioneering efforts at the University of Maryland Baltimore County (UMBC), driven by the Bio-MOD vision, bring us closer to a future where portable, rapid, and tailored pharmaceutical manufacturing can meet the urgent needs of affected populations. This progress represents a significant step towards achieving equitable access to life-saving therapies worldwide.
Shayan Borhani
Doctoral Student, University of Maryland Baltimore County
Shayan is a doctoral student within the Center for Advanced Sensor Technology (CAST) at UMBC. CAST promotes the development of optical-based sensors for bioprocess, environmental and homeland defense applications. For the last seven years, Shayan has worked under the supervision of Dr. Govind Rao on the development of an automated and portable medicines-on-demand device, BioMOD for the small-scale manufacture of therapeutic-grade biologics on a timescale of hours.