What would help you trust novel treatments?

• studies that prove effectiveness
• full disclosure
• open communication
• recommendation by a doctor

The first time we got in touch with scientists working on bacteriophage therapy was during the opening event of the BioKitchen laboratory in Munich. This modern lab offers a well-equipped place for students and start-ups with innovative ideas to perform biotechnological experiments. The central topics of the opening event of the BioKitchen covered the emerging antibiotics crisis and the opportunities of bacteriophage therapy as an alternative treatment. Therefore, attending this event was a major opportunity for us to get in touch with experts working in the field of bacteriophages.

At the opening event we had the opportunity to demonstrate the assembly of our bacteriophages in a hands-on session with other participants. There we had the pleasure to meet phage experts from the Eliava Institute in Georgia (Europe) and Dr. Jean-Paul Pirnay from the Queen Astrid Military Hospital in Belgium. They gave us helpful insights into the practical application of bacteriophage therapy. After fruitful discussions we refocused our project to manufacture therapeutic phages with our affordable cell-free system.

One highlight on our journey was the meeting with several phage experts in Brussels. The goal of this meeting was the creation of a team of scientists and industrial partners to overcome the most recent problems of bacteriophage therapy.

We were invited to build a scientific consortium with:

• Dr. Jean-Paul Pirnay | Queen Astrid Military Hospital Belgium
• Dr. Cedric Lood | KU Leuven
• Prof. Yves Briers | Ghent University
• Dr. Michiel Stock | Ghent University
• Dr. An Van den Bossche | Sciensano

During the meeting we convinced the experts that we can contribute substantially to the safety and efficacy of bacteriophage manufacturing. Furthermore, we learned about Good Manufacturing Practice and the magistral preparation of bacteriophages for therapeutic application in Belgium. To fulfill the legal requirements of a manufacturing process, we streamlined Phactory to comply with the framework of the Good Manufacturing Practice. Moreover, we got the chance to visit their laboratories, where phages for therapeutic use are produced.

Belgium, France and the Netherlands allowed the magistral preparation for phage therapeutics as suggested.

Dr. Jean-Paul Pirnay

It was of great importance to us to demonstrate that medically relevant bacteriophages can be manufactured with Phactory. We received the SEC-3S bacteriophage from the Queen Astrid Hospital as well as the CLB-P2 and CLB-P3 bacteriophages from the group of Laurent Debarbieux at the Pasteur Institute in Paris. These bacteriophages are specific for the E. coli strain O104:H4, also known as EHEC, which caused 53 deaths during an outbreak in Germany in 2011. As indicated in the microscopy images, we successfully assembled these medically relevant bacteriophages in our cell-free system.

We conducted a LAL-test and compared the results with magistral bacteriophage preparation regulations. According to these results, our bacteriophages are orally applicable. If we reach higher titers in Phactory, we can apply extensive purification protocols to allow for intravenous application of bacteriophage therapy.

The advance of the antibiotics crisis in our hometown was of highest interest to us.

To get an overview of the situation, we visited the university hospital Rechts der Isar of the Technical University of Munich to meet Dr. Friedemann Gebhardt, the head of the hygiene department. He is responsible for preventing the spread of drug-resistant bacteria.

Until recently, most of his patients had acquired their multi-resistant infections during travels to developing countries on other continents. This has changed and the problem has spread to Europe. The number of patients with infections caused by bacteria resistant against multiple antibiotics has nearly doubled in the last year, leaving Dr. Gebhardt very concerned.

Dr. Gebhardt showed us around the diagnostic laboratory of the hospital, where patient samples are analyzed. He then brought up a very important fact: To avoid delays in the delivery of life-saving therapeutics, the bacteriophages could be produced directly in the hospital itself. To be suitable for on-site production, sufficient stability of the components for distribution and storage has to be made possible.

Thus, we decided to establish a protocol for lyophilization of the cell-free expression system to fulfill these requirements.