OUR PRODUCT
Viraless is a novel solution to emerging biological threats that rapidly detect the presence of pathogens in different fluids even at the attomolar levels. Fast, accurate, and specific results are obtained through an antibody-based biosensing system on optic fibers. We functionalize optic fibers by attaching one or multiple monoclonal/polyclonal antibodies to their surface. Thus, detection occurs when viral antigens bind to these antibodies and cause a change in the refractive index within the fiber. This interference is then processed through our hardware tool. To make our product more user-friendly, we have supplied Viraless with the necessary software so that anyone can use our tool and obtain already-interpreted results.
APPLICATIONS AND END-USERS
The primary area of Viraless use is mitigation approaches and infrastructure necessary for biosecurity and biodefense. Although there are already many detection methods in place, our novelty is that Viraless can detect synthetic biology-enabled engineered pathogens as well. Since it can be used in its stationary state or as a portable device, it has 4 main applications.
BIOPROCESSING
To upscale the bioprocessing of our recombinant protein system to an industrial level we need to go through upstream and downstream processing steps. For upstream bioprocessing, we will identify the expression rate of recombinant protein using algorithms developed to determine the expression of T7 polymerase. Received data will allow us to calculate the optimal quantity of IPTG in production. However, as we discovered in our project, these proteins will be best expressed in different expression plasmids like pET22b since it contains signal peptide sequences for periplasmic import and could contain a better promoter. It is important to note that recombinant proteins are required in very small quantities only to test the binding capacity of our antibodies on optic fiber (e.g. we can verify that kilometers of optic fiber work as expected using a few microliters of recombinant protein).
We acknowledge that the production of monoclonal/polyclonal antibodies for functionalization on an industrial scale would be expensive. Therefore from a long-term perspective, we plan to functionalize aptamers on optic fibers instead of antibodies because they can be easily made specific for any pathogens, modifiable, and low cost in production. To accomplish this, we built upon existing software and wrote additional tools that allow us to obtain optimal aptamer sequences for our system.
Lastly, the utilization of full-size equipment like LUNA for obtaining readings from optic fiber does not only limit applications of our system, but it is also not affordable and requires advanced training for end-users. Therefore, we scaled down and adapted necessary equipment pieces into a backpack-sized hardware system that can be set up in its stationary form for wastewater systems or used as a portable tool by field epidemiologists.
As a result, we can produce aptamers depending on customers' request pathogen of detection on large scale, supplying kilometers of optic fiber, mass manufacturing our portable hardware, and supplying all end-users with easy-to-use software.
CHALLENGES
The refunctionalization process could be costly and require advanced specialized staff. Not only that but we have yet to determine for how long our system can maintain such high sensitivity rates because as time passes antibodies or aptamers will denature and detach from the surface of optic fiber. This process could be sped up by different toxins and chemicals in environmental samples, limiting the efficiency of our tool in its stationary phase. However, we need only small quantities of liquid for detection (500 microliters), which keeps its portable form usable. At the same time, challenges are expected to arise when it comes to hardware manufacturing on our own, therefore it would be more feasible to buy ready-made tools according to our design from manufacturers that specialize in producing spectrophotometers and other pieces of equipment.
Regarding the commercialization process in itself, we will stick to B2G (business-to-government) model, which could be our advantage yet limitation. Legislative processes of many countries’ defense infrastructure could deny them from utilizing our equipment and/or software. To limit such obstacles we decided not to use cloud-based storage for our software so that it only runs locally, ensuring our end-users that we will not have access to their data. However, we yet have to implement end-to-end encryption into the software.
SAFETY
We have also assessed the safety of our project through the framework for assessing concerns proposed by the National Academy of Sciences, Committee on Strategies for Identifying and Addressing Potential Biodefense Vulnerabilities Posed by Synthetic Biology. Among these, our project could raise concerns in these fields:
Since we work on synthesizing recombinant proteins of known pathogens it could be assumed that we synthesize virulent and dangerous proteins. However, these recombinant proteins are not virulent on their own and cannot be hazardous unless they are a part of fully replicated viral structures (even like that, the system is not likely to work).
As explained earlier, recombinant proteins are 1) safe and non-virulent, 2) are used only in small quantities to verify that our antibody-coated optic-fiber system is capable of binding and detection. None of the used methods or procedures are expected to make bacteria of our interest (E.coli) more pathogenic either.
There is a concern that the easier it is to use the technology, the easier it becomes to deliberately
misuse it for malicious purposes. However, as we reiterated previously, we do not work directly with the
sequence of the pathogens nor pathogens themselves; our end products are neither virulent nor hazardous,
and our hardware cannot be used as a weapon.
As outlined earlier, our project inherently is not capable of producing virulent or pathogenic biological parts. We have followed standard and publicly available molecular biology protocols. We made sure to clearly state everywhere that we are not working with pathogens nor are attempting to create new ones, as well as avoided using an original sequence of the vaccinia virus in our project. We hope to make clear that our goal is to detect biothreats, not create them.