Team:Bielefeld-CeBiTec/HP/Gold Integrated

Integrated Human Practice

Discussion with Cell Product Purification Experts

Feedback and advice from experts actually working in the fields of concern are valuable contributions to a successful iGEM project. To get hints how to improve our purification column EluX, we met with two experts: Prof. Dr. Dirk Lütkemeyer, General Manager of BIBITEC GmbH, and Dr. Benjamin Müller, CEO of Biofidus AG for Analytical Services. BIBITEC GmbH was founded in 2001 and is specialized in the production of recombinant proteins and monoclonal antibodies. Biofidus AG is a bioanalytical company offering services in a wide variety of bioanalytical methods. They are specialized on spectroscopic, chromatographic or mass spectrometric assays focused on characterization of proteins as well as small molecules. We demonstrated and explained our prototypes and elution technique. We also prepared specific questions, like ‘what do you think about our elution technique compared to common techniques?’ and ‘Which problems and limitations could our column encounter in real world applications?’. As described in the following sections, we benefited greatly from these exchanges and made several improvements to our hardware to address the predicted challenges. Therefore, we are very thankful for their opinions and great advices.

Meeting with Prof. Dr. Dirk Lütkemeyer, General Manager of BIBITEC GmbH

We met Prof. Dr. Dirk Lütkemeyer at the 24th of August at the Center for Biotechnology. We presented our purification column and explained our light-induced elution technique. The presentation was followed by a fruitful discussion about possible challenges in real world applications like using this purification technique to purify recombinant proteins produced by our local fermentation technologies working group.

Figure 1: Meeting with Prof. Dr. Lütkemeyer.
Prof. Dr. Dirk Lütkemeyer (BIBITEC GmbH) and team member Yannic during a meeting at the Center for Biotechnology. A detailed discussion provided us with useful advices and opinions about our elution technique and purification column.

Prof. Lütkemeyer stated that our technique of the light-induced elution is very interesting and a promising alternative to proteases-cleaving sites. However, we have to validate the system profoundly. Especially interesting are the minimal amounts of light and radiation time needed for reliable back bone cleavage. He also said that our system could be very useful for companies related to the production of biomolecules like Evonik Industries AG and GE Healthcare. Nevertheless, we should improve our purification column prototype design to reduce dead space, increase potential yield and enhance the scalability. Finally, he recommended to focus on the validation of our elution system, since this is a highly innovative and promising aspect.

Meeting with Dr. Benjamin Müller, CEO of Biofidus AG

We met Dr. Benjamin Müller (CEO of Biofidus AG) on the 6th September at his office (Figure 2). We showed him our peptide purification column and discussed the possibilities and conceivable use cases of our light-induced elution technique. He commented on the advantage of our purification method, namely that the one step reaction is simple and easy to understand and thus attractive for possible users.

Figure 2: Meeting with Dr. Benjamin Müller
Meeting of Yannic Kerkhoff with Dr. Benjamin Müller at his office on the 6th September to discuss the light-induced elution technique.

He also pointed out some things we will have to check if we want to proclaim a real use of this elution technique. He stated that we would have to investigate if the light that is used for the elution will not harm the target protein and if there are any restrictions to the light-spectra, intensity, or irradiation time. We should also validate the stability of the amino acid itself, namely if it only reacts after irradiation, or possibly in the storage process. This would lead to a noticeable loss of the target protein. Furthermore, he stated that it would be useful to create a kit for the light-induced elution so that all users can apply this technique to their desired process and make the usage as easy as possible.

Meeting with Prof. Dr. Thomas Noll and Ole Weigelt

To discuss possible marketing and commercialization strategies for our EluX technology, we were invited to a meeting by Prof. Thomas Noll, co-founder of the local biotech Xell AG, and Ole Weigelt, tax consultant and lawyer at Weigelt Miersbach Uhlemeyer Partnerschaftsgesellschaft. The meeting took place on the 7th of September at Ole Weigelt’s office.

Figure 3: Meeting with Prof. Dr. Thomas Noll and Ole Weigelt
Group picture after a fruitful meeting about possible marketing and commercialization strategies for our EluX technology.From left to right: Yannic Kerkhoff, Prof. Thomas Noll (co-founder of Xell AG), Ole Weigelt (co-founder of Xell AG, lawyer and tax consultant) and Christopher Whitford.

Prof. Noll and Mr. Weigelt both stated that our idea has a high innovation potential and could be used as a new purification method. As examples they named the very fragile factor VIII and protein A and protein G due to their expensive column material. Since the iGEM idea is based on making your research open source and share your results with the whole community, Prof. Noll and Mr. Weigelt said that an intense scientific analyzation of the method is more adequate as commercializing it. Both encouraged us to pursue our idea properly and to continue the work even after the competition either by ourselves or by handing over our results to other students of our university, who could analyze this technique in bachelor or master theses.
Both experts pointed out how multifaceted our concept is, as it includes basic molecular biology, cell development, photometry, linker design, cleavage kinetics and media optimization. Eventually, Mr. Weigelt and Prof. Noll advised us in testing the range of wave lengths and light intensities suitable for the cleavage of the target protein without harming either the target protein or the affinity tag. We thus started some investigations into how the UV-light could potentially harm our proteins.

To be decided

Investigation of Public Reception of Synthetic Biology

In order to assess the public opinion of synthetic biology and synthetic life, as well as thinkable applications and attitudes towards them, we designed an online and print questionnaire to be handed out to students, pupils and the general public over the course of two months. The nine-page questionnaire assessed the following constructs:

      1. Attitudes towards synthetic biology The participant’s attitude towards synthetic biology was assessed with 10 questions (e.g. “The risks synthetic biology harbors are uncontrollable”) using a 6-point Likert-scale [1] ranging from “disagree completely” to “agree completely” with an “I don’t know” option.

      2. Perceived dangers of synthetic biology The perceived dangers of synthetic biology in comparison to possible dangers rooting from other sources were assessed with eight scenarios on a 6-point Likert-scale ranging from “very unsafe” to “very safe” with an “I don’t know” option. Four of those scenarios involved synthetic biology (e.g. “Incorporation of non-natural components into the DNA of a bacterium”), the other four were unrelated (e.g. “A several hours-long commercial flight overseas”).

      4. Previous experiences with synthetic biology The participants’ previous experiences were directly assessed by asking whether or not they have gotten in contact with this topic. If that was the case, they were asked to state the origin of their experiences (e.g. “profession”, “volunteer work”, “politics”).

      5. Definition of life The participants’ definition of “life” was assessed by asking them to choose one out of four different possible definitions of “life”, which were coded philosophical, scientific, religious, or judicial. Afterwards, participants were asked on a five-point scale to what extent the chosen definition fits their personal definition. If the definition did not or did hardly fit, participants were asked to write down their own definition of “life”.

      6. Attitudes towards artificial life The participant’s attitude towards artificial life was assessed with six questions using a 6-point Likert-scale ranging from “disagree completely” to “agree completely” with an “I don’t know” option (e.g. “life is life – it does not matter whether its origin is artificial or natural”)

      7. Evaluation whether certain forms of life are natural or synthetic Participants’ opinion on what is natural and what is synthetic life was assessed by six scenarios (e.g. “a bacterium engineered in the lab”) with the options “natural life” “artificial life”, “no life at all” and “I don’t know”.

      8. General attitudes towards our project After a two-sentence description of our iGEM-project, participants were asked in three questions whether our project was safe, meaningful, and whether or not – if the participant had the opportunity to decide – it could proceed. Options were “yes”, “no”, and “I don’t know”.

      9. Demographics Lastly, participants’ age, gender, education, and profession were assessed to further contextualize results.

Sources: [1] Likert, R. (1932). A technique for the measurement of attitudes. Archives of psychology.

Nagoya-Protocol


The “Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization (ABS) to the Convention on Biological Diversity” entered into force on October 12th, 2014 in Nagoya. It is an international environmental agreement to implement the objectives of the 1993 UN Convention on biological diversity: “the fair and equitable sharing of benefit arising out of the utilization of genetic resources” [2]. Especially developing countries which often have a manifold biodiversity are affected by bio piracy. The Nagoya Protocol intends to ensure that these countries at least profit financially or non-financially by the resulting research and products of their genetic resources. In addition to access to genetic resources, it covers traditional knowledge concerning these resources. Contracting countries are bound to take action to guarantee informed consent prior to any course of action as well as impartial benefit-sharing, respecting local laws as well as customary use and exchange.

Today, 100 countries signed the Nagoya Protocol [3]. To succeed, every partner nation should establish ABS National Focal Points, competent national authorities, legislative administrative, national databases and checkpoints for information.

Germany signed the Nagoya protocol on July 20th, 2016. Our donor of the isoG-metabolism-pathway is Croton tiglium, which is a herb in traditional Asian medicine. Its original occurrence is in the Asian region. To make sure we do not infringe the Nagoya Protocol regulations, we clarified the origin of our plant as we got it from the botanical garden of the Phillips University Marburg. Croton tiglium is originally from an undocumented old stock of the botanical garden Giessen. In 1986, the botanical garden Marburg received seeds from this stock. The date on which the EU regulation entered into force was October 2014. All resources collected before this date are not subject to reporting requirements of the Nagoya Protocol in Germany. Thomas Greiber (Federal Agency for Nature Conservation, Head of Division Div I 1.4 “Competent National Authority for the Nagoya Protocol”) confirmed “[..]Therefore, the respective areas of application are not touched by this EU-regulation, and the mentioned regulations are not relevant.”

Therefore, we are allowed to work with the plant and publish information and results about gene sequences and metabolism pathways without violating applicable law.

Merck

Merck is a leading company for products in the pharmaceutical and chemical sectors. Their expertise consists of three mayor sectors: Healthcare, Life Science and Performance Materials. This wide variety of subject areas ensures Mercks status as a global player. Their close collaboration with scientists ensures the high-quality of their products and know-how. Merck has supported the iGEM Bielefed-CeBiTec teams for seven years. We are grateful for their continues scientific support and expertise. Back in August, we visited Merck to present our final project idea and the first results. Our visit was organized by Prof. Dr. Herget, who has been a great supporter of the iGEM-Teams Bielefeld for several years.

Figure 2: iGEM Bielefeld-CeBiTec 2017 team members Olga Schmidt and Markus Haak presenting our project in front of the Merck scientists.

The following discussion dealt particularly with possible in vitro and in vivo approaches and the ethical concerns regarding our project. They confirmed our idea to add in vitro experiments as an additional control for our project. The moral concers have also been addressed. Therefore we designed a survey and asked for second opinions in a variety of meetings with ethicists. This eventuated in our report “ChImp - Chances and Implication of an Expanded Genetic Code”, from which we derive guidelines and recommendations for work awareness in science. Visiting Merck is valuable in more than just one way: we did not just gain experience in holding a presentation in front of experienced researchers, but also received great advice for the future development of our project. We are excited to visit Merck again next year to present our final results and the progress we have made.

Figure 3: iGEM Bielefeld 2017 team members Olga Schmidt and Markus Haak with representatives from the iGEM Bielefeld 2016 team and Merck.