- The team developed a reproducible biochemical analysis approach for phage genome sequencing using a portable DNA sequencer called the MinION to make the approach affordable and accessible. They successfully tested their approach by sequencing a mono-phage sample from Ukraine and discovered a new phage genome. - Moving forward, they plan to engage students through school projects using their approach, conduct mobile phage sequencing labs in rural areas using the portability of their method, and develop an online phage database to accelerate phage research.

1. Phage4Life
Read Deck

2. Our Team
Introduction
Our team is called Phage4Life and consists of two team members, Philipp and Niklas. We both are Management &
Technology students at the Technical University of Munich in our master's degree and participants in the
TechChallenge 2019/20 at UnternehmerTUM. Until October 2019, we both had absolutely no background or practical
experience in biology and chemistry. However, we were highly motivated to constantly push ourselves to learn something
completely new and different besides our studies. The TechChallenge is an incredible opportunity to achieve this goal as well
as to collaboratively work on real projects and educate ourselves hands-on in practice. This represents a perfect contrast to the
overwhelming theoretical education we receive in university and is very refreshing, interesting and, most of all, it makes fun.
The importance of the antibiotics challenge at the Bio.Kitchen raised our attention as it is a really bad situation for every
human being if nobody is going to find a feasible solution. Multi-resistant bacteria could probably kill millions of people around
the world as none of the existing antibiotics can destroy them anymore. We asked ourselves, if we really want to ignore this
huge problem, and our answer was NO! So finally, we decided to tackle this challenge in the TechChallenge 2019/20 in order to
contribute to this immensely important topic.
Today, we must say that we are incredibly proud of what we achieved so far and think what we discovered could really change
everybody's life for the better. We are Phage4Life, and we’re going to safe your life!
Philipp
Niklas

3. Customer Need (1)
Global Scale
In the modern world, doctors use a lot of antibiotics in order to support the human body to fight against harmful bacteria. This is best practice in the EU since
antibiotics got famous with the use of penicillin in the 1940s. But there is a big problem: Over time, bacteria develop resistances against certain antibiotics. The
consequence is, that such antibiotics become completely useless and therefore can not be further applied in medicine. Although these issues are well known,
pharmacy companies stopped investing in antibiotic research because it is not profitable enough anymore. This means, at a certain point in the future we will
have many multi-resistant bacteria and no antibiotics which can fight them. The problem is, that the resulting multi-resistant bacteria are going to be killer
machines. Researchers predict, that by 2050 more than 10 million people are killed annually due to such bacteria. Considering this information, the need for
an alternative to antibiotics gets obvious. Luckily, there is an alternative, the so-called bacteriophages – or short phages.
A main advantage of phages compared to antibiotics is, that phages are viruses with an average size of 200 nm which are only harmful for bacteria and not for
human body cells. Phages fight bacteria by injecting them their own DNA which forces the bacteria to produce many more new phages until itself explodes.
Although phages were discovered more than 100 years ago, we do not use them in medicine. This has two main reasons. According to the rules of ethic by the
World Medical Association, phages are only allowed to be used when there is no other option available. Secondly, especially during WW2 antibiotics
outperformed phages, because one phage can only eliminate one exact opponent bacteria. In order to use phages here in European medicine, we need to
know exactly which phage can destroy which bacteria. This can be done through comparing the genome of phages to the genome of the bacteria. Most phages
are biologically undiscovered which means we do not know their DNA. Therefore, we have a global need for further research on phages as they are a
possible solution to the antibiotic problem.

4. Customer Need (2)
Local Scale
Within the area of biochemistry, we discovered a general need for reproducible biochemical analysis approaches. Such approaches can be used in large
scales and by many who have access to fully equipped laboratories in order to contribute to a common goal and to scientifically investigate the same subject.
However, the access to fully equipped laboratories is not always the case. Therefore, we also discovered the need for affordable equipment, which is easy to
use, sometimes even portable but which provides the same functionalities as the more expensive professional equipment. Such affordable, simple and
portable equipment can further lever the scaling effects of reproducible biochemical analysis approaches as many more researchers can participate in
the research and can collaboratively investigate the subject. This then not only includes professional researchers but also unprofessional researchers like pupils
in schools or students in universities.
Summary
• The need for
alternatives to
antibiotics
Antibiotic
Crisis
• The need
for more
knowledge
about phages
Phages as
alternative • The need for
acceleration in
phage
sequencing
Research on
phages
• The need for
reproducibility,
affordability,
portability
A sequencing
approach

5. Value Proposition (1)
A reproducible biochemical analysis approach for phage genome sequencing
Based on the mentioned problems like the lack of reproducible biochemical analysis approaches and the unpopular research on phages as an alternative to
antibiotics, we decided to develop a reproducible biochemical analysis approach for phage genome sequencing in order to create publicity and speed for this
important topic. Our approach basically consists of two steps. The first step is the DNA extraction. When having phages in a tube, this tube also includes
interfering substances like proteins which must be purified prior to DNA sequencing. The whole purification steps are explained in detail in our technical project
description. The second step is the actual sequencing of the DNA. It is performed with the help of the so-called MinION from Oxford Nanopore. With a size
of a USB drive, the MinION is the only portable DNA sequencer in the world. As it is available for 1,200 € only, it is relatively affordable compared to
professional sequencing equipment. The price advantage and the portability of the MinION was very important to us. Furthermore, Oxford Nanopore provides
you with a lot of useful and understandable information and step-by-step instructions to get your sequencing tasks done. The use of the MinION in the
sequencing step therefore makes our whole analysis approach reproducible, affordable and portable. The resulting data from the sequencing step is
afterwards bioinformatically interpreted. We finally came up with the full genome of a new phage.

6. Value Proposition (2)
Proof of concept and the discovery of a new phage
Phages are everywhere, especially at places where lots of bacteria live. In order to test our analysis approach at the end, we had to think of where we get a
phage from. It turned out that we had the option to collect a sample of water from a water treatment plant and extract a phage from the sample. As this process
is very complex and takes a very long time, we needed another option to get a single phage for testing our analysis approach. Luckily, we were able to
contact the startup Phage24 from Austria which is specialized on phage trades of so-called phage cocktails. Phage cocktails contain hundreds of thousands
of phages in one sample. This means one would still have the work of extracting one single phage before being able to analyze it, so this option wouldn’t really
save time. After discussing this issue with Phage24, they agreed on sending us a sample of a single mono-phage which was exactly what we needed. This
phage was cultivated for our purposes only which is very special and further enabled us to directly start with the sequencing step. We could therefore prove our
approach and its usability. The results were really good and after adjusting some minor changes we finalized our approach in the technical project description.
Summary
• We bought the
portable DNA
Sequencer
MinION
Procuring
MinION
• We got a
mono-phage
from the
Ukraine
Procuring
Phage • We extracted
the Phage
DNA and
purified it
Extracting
Phage DNA
• We performed
the
sequencing on
the MinION
Sequencing
Phage DNA • We discovered
a new genome
and proved
our concept
Discovery

7. Our Achievements
The MinION from Oxford Nanopore
We successfully bought a professional device for our project. According to Oxford Nanopore, the MinION is the only portable DNA
sequencer in the world. We needed it for the phage genome sequencing as our goal was to develop a simple and reproducible
approach which can executed everywhere. For this purpose, the MinION was by far the best decision.
A successful crowdfunding campaign
During planning this project, we faced a financing issue as the portable MinION from Oxford Nanopore costs about 1,200 €. Since
our budget was limited to 250 €, we had to come up with an idea, and so we did. In mid-November we launched our own
crowdfunding campaign on Startnext.com! We proudly raised 152% of our financing goal in exactly one month, which is equivalent
to 610 €. Subtracting the fees, we finally received more than 500 €. Due to this incredible support from the society, we know that
the antibiotic challenge is generally seen as a problem worth solving and that the value created by our project is recognized.
The Mono-Phage from the Ukraine
We can proudly say that researchers from Ukraine supported our project and agreed on cultivating a new Phage for our purposes
only. This phage was the first one sequenced with our approach which is a huge achievement for us.

8. Future Roadmap
We have multiple ideas in mind what can be done and shown by the success of our TechChallenge project. Every idea aims on the further enhancing phage
research and solving the described customer needs.
School projects on phage genome sequencing
This idea is our favorite. When we can learn how to sequence phages, high schoolers will be able as well! Since the approach can be easily learned, it could be
an interesting project in schools or small university courses. The device is affordable and the laboratory equipment we needed will be available in every
standard laboratory. This could not only enhance research on this topic, but as well motivate young and future scientists to engage with phages.
Mobile labs in rural areas
Pack our stuff and off we go! Since only a few things are needed, we can go into the field and do it wherever we want to. The MinION is portable, and the
approach can therefore be done in every area. Some steps will be tricky without a laboratory, but it is feasible.
A phage database
The database plan is still in our minds. Building up one can still be a reasonable solution as a foundation for further phage research. This can also accelerate
the use of phages in modern medicine.