It took Stellenbosch University Ph.D. graduates Du Preez van Staden and Ross Vermeulen seventeen years and more than 24 international papers later to develop their unique skills and technology in heterologous protein production and purification to supply the local South African market with proteins or enzymes of interest.
As the founders of SU and Innovus’s latest spinout company — FluoroBiotech — these two molecular biologists say heterologous* protein production is not a "new" invention, but their technology challenges the protein expression norms and makes it more accessible to the emerging South. Scientists use heterologous protein production worldwide to produce recombinant proteins; these include specialized proteins and enzymes used in basic research, pharmaceuticals (such as insulin), and diagnostics (like the COVID-19 test). But there is a desperate need for locally produced recombinant proteins as most are currently imported. Their vision is to produce proteins or enzymes locally in more significant quantities and on demand so that local biotechnology ventures are supported - and they are well on their way to fulfilling this dream. In FluoroBiotech's lab, currently in Stellenbosch University's Department of Microbiology, molecular biologists Van Staden and Vermeulen spend much time – like other researchers around the globe – trying to solve biological problems. The subject of their research was anti-listeria peptides, small proteins that killed Listeria. "Our biggest problem was not how to treat Listeria, but how hard it was to produce enough of this peptide to understand how it could kill Listeria so effectively. We soon realised that there are always two aspects at play during biological research: one part of the work, the romantic side, revolves around testing a compound to measure how effective and safe it is,” says Vermeulen. “Fighting, for example, antibiotic resistance a cure for cancer, high blood pressure, or Alzheimer's treatments are all revolutionary and virtuous endeavours. However, a researcher must first gain access to enough of a compound to be tested; this side of research does not often find the limelight. To this, many of the most interesting compounds that need to be tested are proteins that one cannot buy commercially, so they must be manufactured.” Coined "iteration" hell – was the place younger Van Staden and Vermeulen soon found themselves in with the production of their anti-listerial peptide. Except, they did not have access to a first-world budget or equipment, which meant there was no room for blindly iterating the expression system. Production needed to move forward with severe intent, which meant challenging every established production and iteration protocol. "Initially, we were interested in peptide antibiotics which were very effective against Listeria and could be used to prevent its growth. However, before we could start to test how effective and stable these peptides were, we needed to produce them. This took us down a rabbit hole of protein expression endeavours, which has ultimately allowed us access to many different proteins with a much wider range of biological activities, not just ant-listeria activity," said Vermeulen "Heterologous expression* started as a monumental challenge with limited resources and developed into a passion and a point of pride. We have significantly improved the robustness and efficiency of producing heterologous proteins in the bacteria known as E. coli. This is because necessity is the mother of invention, and for our research, we needed to produce proteins faster and cheaper – routinely."
The duo realised that protein production is a significant and common problem for scientists and often causes major delays and deviation in related biological research. In our opinion, this is a severe hindrance in third-world countries that cannot outsource the work due to the associated costs. "We realised that over the years, we had created a precious heterologous protein expression system that can work better to produce many already valued proteins because it fundamentally consolidates the production process. The great thing about this is that you will probably find an enzyme that Mother Nature has produced to solve any biologically related problem," said Vermeulen
"The fact that there is life on earth means that Mother Nature has overcome a monumental number of obstacles/challenges/problems to arrive at this point. It has done this by creating proteins or enzymes, each having a function in a given biological system. Enzymes are the fundamental driver of life. Biologists like to use the term catalyst because they speed up a certain process, but you can think of them as extremely tiny problem solvers working in a microscopic factory which is a cell."
Vermeulen said that because humans are made of, and by, proteins, many of humanity's problems can be solved using enzymes and proteins. Still the trick is to produce them in significant quantities correctly. "There are countless ways proteins are applied in society; the incredible part is that more are being discovered daily. Massive research efforts are dedicated to prospecting new enzymes. This prospecting occurs because the information to manufacture a protein inside a cell is stored as sequential information in a DNA molecule. Currently, massive amounts of DNA are being sequenced worldwide in the process of next-generation sequencing, supplying an endless number of protein "blueprints".
FluoroBiotech's specialty is to put these blueprints into microbial cells, which are very good at converting the DNA information into large amounts of a functional protein (when genetically modified). This process is known as heterologous expression and is used worldwide to produce large amounts of target proteins. "Overseas, researchers like us often have lots more money, ample facilities, and no load shedding. We also import most of our recombinant enzymes at a hefty premium to perform our work. Research often feels like you are shoveling against the tide, so these additional constraints mean things are done differently here". It was during the COVID-19 pandemic that the duo realised just how severe these constraints were for the country, especially in terms of enzyme imports. During the pandemic, all enzyme imports were halted, and it became evident how desperately South Africa needed a local enzyme and protein supplier or specialist.
"The solution to some of these problems was sitting in our freezer (we store genetically modified organisms carrying our biotechnologies in the freezer)." “During the first hard lockdown, while helping a biotechnology company, I realized that our expertise and technology have a commercial application and the potential to help many people. It was also when imports into South Africa were restricted, limiting research activities for academia and industry, making the need for a company like Fluorobiotech painfully obvious. I then knocked on Innovus's door and pitched the idea of commercialising our services and technologies. They were hooked and asked us to start building. From here, we refined our protein production platform, and with the support of the teams at Innovus and LaunchLab we got Fluorobiotech off the ground,” said Van Staden. Much engineering and ingenuity went into FluoroBiotech's system to make it as robust and efficient as possible because it had to accommodate many more challenges like load-shedding. Currently, they serve smaller clients, such as university scientists who are trying to solve big biological problems and need specific proteins or enzymes, but this is just the beginning. Their dream is enormous, and they believe FluoroBiotech could be the company that will kickstart the biotech industry of South Africa because proteins are the bread and butter of the international biotech market. They also are confident that their simplified E. coli expression system, which produces more protein with less effort and resources, will be globally adopted. The proof is in the pudding and these "molecular chefs" cook on all burners.Having secured their first investment from the University Technology Fund, with the help of Innovus, Vermeulen and Van Staden are confident they are building a sustainable business. "The competitiveness of many technologies can in some ways be compared to a sports match - it's often the fitter team that will win: the team or technology that can do more with less: time, resources, opportunities, or downright luck."
Vermeulen says South African research in the biological field is buzzing because South Africa faces the same biological problems (healthcare, agriculture and climate change) as other countries. Still, we are trying to solve them against all additional odds. "We believe that the local ingenuity, drive, and talent are just waiting to be released on the global biotech market - but it needs essential services like protein production. FluoroBiotech wants to catalyse this by supplying proteins locally and at more competitive prices using our expression tech. For this reason, we are now setting our sites on our production lines and constructing a local "stadium" per se – a commercialization laboratory to marry R&D with hopefully GMP to sell proteins and biotechnological solutions to a global audience," said Vermeulen
Vermeulen and Van Staden believe that if they can roll out their technology locally, it could meet the demand and democratize the current protein expression techniques to some degree. South Africa can begin producing heterologous proteins and solutions for export. This is a space to watch.
* Heterologous expression refers to the expression of a gene or part of a gene in a host organism, called the heterologous host (like E. coli), that does not naturally have the gene or gene fragment in question. Insertion of the gene in the heterologous host is performed by recombinant DNA technology. For example, you can put a human gene (like the gene that codes for insulin) into a bacteria and make that protein inside the bacteria.
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