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Since the beginning of March, Applikon Biotechnology has been working with the Laboratory of Virology and the Bioprocess Engineering Group of Wageningen University on the development of a COVID-19 vaccine, based on mimicking the virus’s “spike protein,” according to Arthur Oudshoorn, the CEO of the company, which is a subsidiary of Getinge. The virus uses the protein to attach to receptors and enter human cells.
The team is using the Sf9 baculovirus-insect cell system to produce the protein for vaccine and serological test applications, adds Oudshoorn.
“The baculovirus expression system is commonly used and has proven to be effective in a number of influenza-type applications,” he says. “It turned out that the pH range for optimal growth of the spike protein is far from what we are used to seeing in cell lines. Insect-cell-type systems do much better in slightly acidic conditions, and we were able to grow a much higher density of cells than was normally the case.”
To produce sufficient amounts of spike protein in a relatively short period of time, rapid screening of cell lines for optimal process conditions is of utmost importance. Applikon’s micro-Matrix instrument is used to screen different conditions for the best possible cell growth and protein production in Sf9 cells,” notes Erik Kakes, sales and marketing director.
“The micro-Matrix system allows for simultaneously performing 24 cultivations in a deep well plate with individual measurement and control of DO, pH, and temperature. For example, screening for optimal pH value can be done in a single experiment using 24 different pH values,” he says. “We do the actual experiments in our facility with our equipment. We use all this information in the scale-up train starting from the micro level. We then go to lab scale, and then we move into larger scale production.”
“We provided the instrumentation that was needed and even the lab space. Otherwise, it would take longer to move forward with the work on the vaccine,” points out Oudshoorn.
Optimal conditions are also checked in Applikon’s 500 mL miniBio (autoclavable bioreactor) and its 500 mL AppliFlex ST (single-use bioreactor) and will eventually be scaled up to a 20 L bioreactor, producing sufficient material for preclinical animal trials and serological testing, continues Kakes.
“MiniBio bioreactors are a true scale down of the laboratory-scale bioreactor,” says Kakes. “These systems can be customized to fit the demands of any process. The small volume reduces media costs and maximizes bench space, which is normally at a premium. Set up and operation are easy, and more data can be generated in less time.”
The AppliFlex ST single-use mini bioreactor provides the same flexibility of the company’s customizable range of glass and stainless-steel bioreactor systems, according to Kakes, who adds that the bioreactor is delivered presterilized and integrates with software automation and automated sampling tools.
“By using 3D printing technology, we can create any bioreactor configuration that optimally accommodates a particular process’s requirements,” he explains.
Oudshoorn is proud of how closely Applikon is collaborating with the researchers at Wageningen University.
“It’s quite unique that an instrumentation manufacturer works with a university like this on a project like this. Normally you are limited to being a supplier. But in this case, a pandemic, it’s on a much different level,” he says. “It’s also necessary to do these things specifically at this time because although we are dealing with a relatively straightforward disease, it is a new viral infection.
“But this is something that’s similar to what the world has seen a number of times in the past. Big Pharma, however, has neglected these infectious diseases simply because the treatment has to be inexpensive. Not enough revenue is involved for the big pharmaceutical companies to jump in on this. Now the whole world is faced with the problem, and a lot of money has suddenly become available.”
Oudshoorn emphasizes the point that the driver behind the partnership with the university is optimizing processes, including scaleup, and making sure that the spike protein yield that is obtained from the process is high because that pushes the cost for treatment down dramatically. “And that’s exactly what we are doing now with Wageningen University,” he says.
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