Day 3

„Made by Synthetic Biology“ – A Status Quo Analysis

Session host Birgit Wiltschi, Head of the Synthetic Biology unit at acib, welcomes the first speaker John Cumbers who made a career at NASA working on extremophile microorganisms, synthetic biology and sustainability. John mentions that “2% of the US-GDP is coming from genetically modified products” and quotes famous Craig Venter who said: “Over the next 20 years Synthetic Biology will become standard for anything”. The costs of reading and writing DNA is decreasing considerably and drives the biotech-industry. Synthetic biology will be used to make biology much easier to engineer. John compares the boom in the computer industry to the upcoming synthetic biology boom. “It’s propelling that a lot of former tech entrepreneurs are now re-investing in the synthetic biology field” as e.g. Bill Gates. Talking about “Cloud labs”, he also introduced several machine-learning processes for microbial strain engineering using dinoflagellates, which keep naturally glowing on your bedside table for about 3 months. John quotes Emily Leproust from Twist Bioscience: “In the not-too-distant future we’ll be able to affordably make bacteria from scratch.”

Mark Bruyninx from Twist Bioscience stepped in as a surprise speaker. Twist, a biotech-start-up, has developed a high-throughput synthesis of DNA to tackle the problems of mankind. “The population is rapidly growing, but resources are finite”, he said and oil-based products must be replaced by sustainable bio-products produced by bio-factories. Also, the amount of arable land is shrinking and there is a need to improve the efficacy of food production, which can be greatly improved by synthetic biology. The company is also working on DNA as a data storage. The amount of data generated is doubling every year and data storage takes up a lot of space and a lot of energy for cooling the servers. Since nature uses DNA for information storage for billions of years and the total content of Wikipedia could be easily stored in a 1,5 mL reaction tube, Microsoft stepped in. Microsoft has purchased 10 million oligonucleotides for digital data storage and reached a byte recovery rate of 100%. In another cooperation Twist and Microsoft stored all music e.g. “Smoke on the Water” using DNA and recovered it later flawlessly.

The next speaker Clemens Lakner from Merck Ventures gave an insight on synthetic biology from the investor site. He admits that venture capital money is expensive, as it expects a high return-ratio and might not fit every business. However, VC-funds invest at a very early stage and a lot of these companies fail so the high return in case of successful companies must compensate these losses. As mentioned earlier there are a lot of tech-investors moving into synthetic biology. “Over the past 20 years the ability to engineer biology has improved dramatically” said Lakner, quoting Drew Endy: “We need to re-write the living world so that it becomes model-able.” He notes that the regulatory framework is still challenging, as is consumer acceptance and at the end quotes researcher Jay Keasling “We have as much to do on the public side as we do on the engineering side”.

The last speaker Harald Koenig, Senior Scientist at the Institute for Technology Assessment and Systems Analysis in Karlsruhe) focuses on the societal, political and ethical aspects of synthetic biology. Harald states that there are already a multitude of reports on synthetic biology e.g. by ethics councils, scientific organization, policy advice institutions, civil society organizations. The consensus from these reports was to avoid hypes and to responsibly use the advances in the field. Biosafety (health, environment) is a big issue and in the future risk assessment may no longer be based on similarities/differences to donor or recipient organisms as it is now. There are also regulatory challenges by genome editing techniques e.g. small genome edits, which could also happen naturally and it’s the question if these changes are then resulting in a GMO or not. Gene drive, being able to spread certain genes rapidly in a given population is also an issue for biosecurity in case of malicious use. The Nagoya protocol was also mentioned as the equitable sharing of benefits from genetic resources is an important issue.

Challenges in design and production of new and emerging vaccines

This session was kicked off with the question “Which factors drive the development of novel vaccines?” discussed in a presentation by Otfried Kistner, Senior Consultant and Independent Vaccine Expert. The answer is the classical “the supply is determined by the demand”, whereby the supply is the medical need as well as the global burden of a disease. However, considering, that the average development time for a vaccine is ten to over 30 years, a rapid response to the market is hardly easy. Typically, the costs for the development of a novel vaccine are around 300 to 800 million dollars. Whereby the major cost drivers are the clinical studies, which are already preceded by non-clinical animal studies and pre-clinical safety studies.
The challenge in developing vaccines often lies within the inconsistency of regulatory organs across the globe, with each facility having their own regulations and requirements. Additionally, the topic vaccination can be an ethical, political and social issue. Besides these bureaucratic and non-scientific hurdles the scientific challenges of the last 20 years, were related to viruses thought to be eradicated as well as the rapid emerge of novel ones. Most of the time conversional vaccine development strategies are not fast enough for such emergency situations. One solution to this problem can be public-private-partnerships, which allow for long-sighted research irrespective of the current demand. This gives us the advantage of “being prepared”.

Reinhard Kirnbauer, a professor of dermatology in Vienna, presented his research in the field of novel vaccine development, in particular broad-spectrum protection against human papillomavirus(HPV)-induced cancer. Till now the typical antigens in licensed HPV vaccines were the major capsid protein L1, which awards protection only against selected HPV types, or the minor capsid protein L2, which induces low-level immunity against type-common epitops. He and his team created a novel vaccine, where the targeted antigen is the chimeric RG1-VLP, a fusion protein between parts of the capisd proteins L1 and L2. This next-generation vaccine induces a broad-spectrum immunity against all relevant mucosal and cutaneous HPV types. For this vaccine, clinical trials are planned for 2019.

Matthias Müller, Head of CMC of Themis Bioscience GmbH, focused on the challenges that go hand in hand with vaccine development for rapidly emerging novel diseases. His answer to the question “How can we develop vaccines against new emerging diseases in an efficient way?” is Themis Bioscience’s novel vaccine platform technology. It can be explained as a “plug and play system”, where a commercial measles vaccine vector and a selected antigen are used to produce a “recombinant vaccine”. As a proof of concept, he presented two of their success stories in novel vaccine development, the MV-CHIK, a vaccine against the Chikungunya disease, and MV-ZIKA, vaccine against the ZIKA virus. One of the main advantages of their technology is the speed with which novel vaccines can be produced – development time can be reduced to as little as twelve months. The discussion was concluded with the question “Is being fast sufficient? … Alternatively we could also be prepared”. Falling into line with Otfried Kistner, Matthias Müller again underlined the importance of vaccine development for cases where there is no market yet.

The session was concluded with a presentation by Udo Reichl, a professor of bioprocess engineering at the University of Magdeburg, who talked about process options and optimization possibilities for high yield viral vaccine production. Conventionally, vaccine production does not happen at high cell densities and 99% is done in batch culture. To answer the question “Are there options to improve the existing production processes?” he presented data of vaccine production in high cell density cultivations. In these fed-batch systems a continuous feeding of the cells is possible, which can potentially lead to 10-100-fold higher product yields. In particular, he introduced the idea of vaccine production by cell cultivation in a tubular plug flow bioreactor (PFBR). As an important fact to be considered he also talked about the importance of process monitoring, which is leading in the direction of quality by design for vaccine production.

Continuous Production

Continuous production of biopharmaceuticals is a current challenge in bioprocess engineering. Especially in downstream processing continuous processes are still in their infancy. In this session chaired by Alois Jungbauer delegates from of academia, industry and a regulatory authority presented their view on continuous processes, especially in downstream processing.

Suzanne Farid, Professor of Bioprocess Systems Engineering from University College London, gave an insight into economic considerations that have to be taken into account when developing production processes and showed various examples from industry. Economic evaluation can be a valuable tool to find the most efficient and cost-effective unit operations for continuous processes.

Gorazd Hribar from the biopharmaceutical company Lek in Slovenia, part of the Novartis group and active in biosimilars production, showed industrial aspects of continuous downstream processing. After introducing several strategies for manufacturing of biologics in general, he showed results obtained in the frame of the H2020 research project NextBiopharmDSP, which aims at implementing a fully integrated manufacturing platform for a biosimilar MAb. The transition from current downstream processing standards to an integrated process could be achieved by implementation of continuous processing, single use operation units and out-scaling instead of up-scaling. Such processes in smaller facilities will have benefits in terms of cost efficiency and environment by saving resources.

Christoph Mück from the Austrian Agency for Health and Food Safety (AGES) presented his view on the regulatory framework regarding continuous production of biopharmaceuticals. Robustness and stability of processes is crucial for the establishment of continuous processes and it is highly recommended to follow a Quality by Design (QbD) approach based on product and process understanding. It is highly recommended to implement the concepts from the ICH guidelines (International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use) which are fundamental for the development of emerging technologies. In principle the current regulatory guidelines enable the implementation of continuous manufacturing processes. An important issue for continuous production processes is to find a suitable definition of a batch.

Scientific Storytelling

In the Special Session “Scientific Storytelling” with Rafael E. Luna, executive Director of the National Research Mentoring Network, Principal Investigator of the Administrative Core of NRMN at Boston College, and author of The Art of Scientific Storytelling, it was pointed out, that storytelling for scientists is getting more and more important. Being published and getting accepted with research results in very early stages of a career is crucial. Scientific journals, on the other side, have lots of papers to choose from. Finding the plot and the right words that “sell” is fundamental. Scientists should prepare for that.

Luna spoke about this topic as an expert in both: He is a high-level researcher and a publicist at the same time. With scientific storytelling he managed to get his papers accepted by the most recommended journals around the globe. It all begins with the title: Every title of a groundbreaking paper has to offer a story with and within the research results granted. With this one sentence of the title, a story has to be worded. A story usually consists of a protagonist, an opponent, a conflict, the solution and the staging where all of this takes place. Luna developed these five parts very comprehensibly out of a common movie plot like the one from The Lion King.

Drawing parallels from the lifecycle in normal research projects and the dramatic arc of a good storyline, the paper itself could be written, according to an arc, such as: introduction, results one to three up to the result climax, the results validation and the closing discussion.

The participants at the workshop were stunned at the easy steps Rafael E. Luna presented. Example titles of successfully accepted papers led to a lively discussion. Thinking of your own research results in terms of a movie could, indeed, add liveliness to scientific writing.

Closing Lecture: Bridging tomorrow

The closing lecture of ESIB2017 deals with biopharmaceutical protein glycosylation and its customization. According to speaker Nico Callewaert from University of Ghent the market for biopharmaceuticals is currently estimated at 250 billion US dollars per year. 60% of these products contain sugars in their structure, such as glycosylases.

The methods for improving the functionality of those glycans are for example “GlycoDelete” technologies that aim for keeping glycosylation as simple as possible. This is highly relevant in the production of vaccinations: By application of “GlycoDelete” the injection frequency of vaccines can be significantly reduced, because the active ingredients are more stable and remain in the blood stream for a longer time.
Another methodology, called “GlycoSwitch” was applied to the B-cellizing in Pichia pastoris: “In yeast production systems, which are used for insulin or also hepatitis B vaccine production, the high level of mannose-glycosylation is a problem”, Callewaert stated. By changing the glycosylation structure and optimizing the tuning pathway, the problem could be solved and, subsequently, Pichia is applicable also to hIgG and Fc fusions, which represent a major part in antibody production.

An important outlook to the future concerning biopharmaceuticals is for Callewaert: “We now have antibodies to respond to diseases, but we are not fast enough to produce them – we have to speed up!” and he already offers possible solutions: “Speeding up could be realized by either rapid lean manufacturing using cell banks of Pichia (ready-to-go semi-continuous production plants) or by neutralizing antibodies that are stockpiled long term in soy bean seeds.”
The vision for the future is to produce the target antibodies within 2 to 3 weeks. As soon as we are able to meet this demand, vaccines cannot only be produced but also arrive early enough to treat our diseases.