Attributions

Science:

• Dr. David Foschepoth for his advice throughout the project and critical questions during presentations
  • Xiabo Zhong, MSc. for his input on plasmids suitable for co-expression of proteins

• Dr. Chao Du for his help in optimizing PCR conditions
  • Camillo Iacometti, MSc. for explaining to us the three different levels of protein expression and possible plasmid designs to utilize for protein expression.
    • Dr. Lennart Schada von Borzyskowski for providing us with the ASKA and KEIO strains, pET16b, and Top10.
      • Jo-Anne Verschoor, MSc. for advise on how to get our DNA constructs into pJET1.2 blunt
        • Dr. Erik Vijgenboom for his input on plasmids suitable for protein expression in E. coli and his guidance on how to best clone our constructs into our plasmid of choice.
          • Prof. Dr. Dennis Claessen and Maarten Lubbers, MSc. for providing us with extra laboratory space after completion of the allocated 2-months lab time during the summer.

Lab Practicalities:

• Ellen de Waal, MSc. and Helga van der Heul, MSc. for assisting us with ordering lab supplies for our project.

• Dr. Lennart Schada von Borzyskowski, Carola Feijt, and Mark Arentshorst for providing us with a plate reader and instructions on how to use it. Jo-Anne Verschoor, MSc., Noortje Dannenberg, MSc. and Camillo Iacometti, MSc. for assisting us with operating the plate reader.

• Dr. Lennart Schada von Borzyskowski for access to a sonicator. Bas van Woudenberg, MSc. and Tijn Delzenne, MSc. for providing instructions on how to use it.

• Kees Koops, the Claessen and Gilles Lab for providing us with a laboratory for our experiments over the summer.

• Hein van der Meer for all the cheerful greetings at the Institute of Biology Leiden every day and occasional ice-cream treats.

• Dame Lab at the Leiden institute of chemistry for assisting us with the sucrose centrifugation gradient protocol and allowing us to use the ultracentrifuge.

• Gerda Lamers and Dr. Joost Willemse for spending hours behind the Transmission Electron Microscope to help us image the nanoparticles.

• Prof. Dr. Michael Richardson and Meiru Wang, MSc. for their insights on cytotoxicity of nanoparticles in chicken embryos.

• Marc Fluttert and Dr. Mark Begieneman for their advice on safety regulations and safety assessment on this project. Also, for their advice and insight on the iGEM safety sheets.

• Dr. Rachel Doherty for allowing us to use the Zetasizer and providing instructions on how to operate it.

• Dr. Agnese Amati for assisting us with laser experiments that were vital to the proof of concept

Outreach:

• Annette Heijn, Corinda van Bohemen, Didi Spaans for assistance with crowdfunding.
  • Didi Spaans for helping us with the audio for our promotion and crowdfunding video.

• Anna Lee for creative insight and filming of our promotion video.
  • Gorlaeus building for letting us use their facilities for our promotion video.
  • Science library for letting us use their facilities for our crowdfunding video.
    • Pim Rusch for making our team pictures.

• For our general outreach we would like to thank all the organizations that promoted us, helped organize events and provided a working space: Nemo Kennislink Biotechnologie, LINDA.meiden, SynBioNL, Leidse Biologen Club (LBC), S.V LIFE, Leiden University, LUMC, PLNT & PLNT Student, unlock_, OVBSP, Science meets Business, Oncode Institute, MARE, Hortus Botanicus, Leiden City of Science, Rijksmuseum Boerhaave, Bioscience Park, Leidsch Dagblad, Leiden Science Magazine and KNCV.

Human Practices:

• Dr. Dick Sterenborg (Universiteit van Amsterdam) for sharing his expertise on laser-mediated therapies.

• Joan Birkhoff (Antoni van Leeuwenhoek hospital) for sharing her knowledge of Photodynamic therapy (PDT).

• Dr. Kristina Djanashvili for teaching us more about imaging techniques, coatings and targeting.

• Dr. Anke Hövels (KWF) for her knowledge of rare cancers, process of treatment development and economical insights.
  • Sofia Dimitriadou, MSc. (Vsparticle) for giving an insight into their methods.

• Dr. Remco de Bree (UMC) for sharing his knowledge of head and neck cancers.
  • Dr. Sylvia van Egmond (LUMC) and Dr. Martin de Jong for sharing their knowledge of current therapies.
    • Dr. Sylvestre Bonnet for sharing his knowledge of nanoparticles and aiding us in experiment design.
      • Dr. Dimitri Pappaioannou (Oncolines) for his knowledge on requirements for clinical trials and targeting.
      • Wessel Teunissen, MSc. and Dr. Cécile van der Vlught-Bergmans (RIVM) for guiding us to safe practice.
        • Dr. Aquiles Carattino (Dispertech) for showing their office and sharing his knowledge on nanoparticles.
        • Dr. Henk Noorman (TU Delft) for his knowledge of upscaling and sustainability in biotech.
        • Lian van Amerongen, MSc. (Medical Delta) for connecting us to practitioners and advising us.

• Aaike van Vught, MSc. (Vsparticle) for his expertise in entrepreneurship and the nanoparticle market.

• Paula van Rossum, MSc. (Toxys) for her expertise on genotoxicity.
  • Dr. Geert Frederix for his insights into economic evaluations and health insurance coverage.
    • Aisha Ahmed, MSc. (LUMC) for sharing her expertise in product design in clinical pharmacology and dealing with the EMA.

Entrepreneurship:

• Stefan Ellenbroek, MSc. (Unlock) for his insights in entrepreneuring in biopharmaceuticals.
• Eugene Golov (Upstream Foods) for his experiences and advice as a former iGEM participant.
• Max Green (Panorama Laboratories) for his entrepreneurial knowledge and his experience with electrochemical synthesis.
• Prof. Dr. Adam Cohen (CHDR) for his inexhaustible knowledge about the pharmaceutical world and for his amusing stories.
• Paula van Rossum, MSc. (Toxys) for her knowledge about genotoxicity.
• Dr. Lorenzo Bombardelli (LURIS) for his advice concerning Intellectual Property.
• Prof. Dr. Henk Noorman (TU Delft) for his knowledge of upscaling and sustainability in biotech.
• Dr. Anke Hövels (KWF) for making us familiar with KWF’s approach.
• Marjolein Crooijmans, MSc. for advising us on the wiki page.

Public Engagement and Education:

• Amber Verhaar, MSc. (Leiden University) for coordinating the Leiden Science Family day
  • Mirjam Stolker (Hortus Leiden) for helping with midzomernachtfestival
    • Chris Jaeger from Leiden City of Science 2022 for helping coordinate our involvement at Leidsch Bierfestival
      • Roos Janssen from the Leids Bierfestival for hosting us there
        • Daniëlle Snijders from the Leidse Biologen Club for hosting us at their Symposium
          • Ellen Zwiers from studyasociation LIFE for having us for a lunch lecture
            • Anniek Mueller for helping and hosting us at Rijksmuseum Boerhaave during the Museum night Leiden.
              • Nathalie Winkster, MSc. for publishing and giving feedback on the NEMO blogs our team wrote.
                • Nicole van 't Wout, MSc. for publishing and giving feedback on the NEMO blogs our team wrote.

    Programming:

    • iGEM Team 2021 from Wageningen University & Research for the coding guidelines
      • Ehsan Razaghi Siahroudi for his suggestions on programming.

    Modeling:

    • Dr. Frans Rodenburg for helping with the statistical analysis of the model.
    • Dr Kemal Sumser for his help modeling in Sim4Life.

    Our contribution:

    Experimental work:

    Name	Project Part	Task description
    Jennifer Adami	Experimental design, plasmid construction and testing WT, transformed, ASKA and KEIO bacterial strains.	Assisted with designing the experimental workflow for the project and constructing the plasmids with genes of interest. Carried out nanoparticle synthesis experiments with transformed strains and selected ASKA and KEIO strains.
    Maxim Velli	Experimental design & chemical synthesis	Designed the experimental workflow, designed engineering cycle, produced bimetallic nanoparticles with chemical synthesis used for proof of concept.
    Jean Richard Quant	Plasmid assembly, bacterial transformation, and modeling experiments	Constructed plasmids with genes of interest and transformed bacterial cells with the plasmids. Assisted with obtaining data relevant to the project model.
    Moor de Waal	Chemical synthesis and general laboratory assistance	Involved with chemical synthesis of bimetallic nanoparticles and assisted with various experimental tasks in the lab.
    Windi Putri Wulandari	Assistance with biological synthesis of nanoparticles and general laboratory assistance	Assisted several team members with various lab experiments linked to biological synthesis, and largely assisted with experimental preparation.
    Olivier Muzerie	Zetasizer experiments and nanoparticle quantification	Assisted with obtaining the hydrodynamic radius of the biologically produced NPs using the Zetasizer. Also, worked on an ideal approach to quantify the number of NPs produced.
    Zainab Rashid	Testing WT, transformed, ASKA and KEIO bacterial strains, and optimal experimental set up for nanoparticle production.	Carried out nanoparticle synthesis experiments with the transformed strains and selected ASKA and KEIO strains. Assisted with identifying a suitable experimental set up for nanoparticle production.
    Eva Pakvis	Chemical synthesis and laser experiments	Produced bimetallic nanoparticles using a chemical method for the proof of concept. Assisted with showing that biologically produced nanoparticles are capable of absorbing laser light.
    Mirthe Zandbelt	TEM imaging, general laboratory assistance	Visualized nanoparticles with TEM under supervision and assisted with various experimental tasks in the lab.
    Nadine van de Brug	Zetasizer experiments, laser experiments and general laboratory assistance.	Conducted experiments with the Zetasizer to obtain the hydrodynamic radius of the biologically produced NPs. Assisted with showing that biologically produced nanoparticles are capable of absorbing laser light. Assisted with various experimental tasks in the lab.
    Elise Stengs	Optimal experimental set up for nanoparticle production, TEM sample preparation	Worked to identify a suitable experimental set up for nanoparticle production. Prepared samples to be imaged by the TEM.
    Tino Hoeksma	Nanoparticle extraction, optimizing conditions for nanoparticle production and production upscaling	Assisted with the extraction protocol of nanoparticles based on size using the sucrose centrifugation gradient. Conducted experiments to identify whether different shake speeds impact nanoparticle morphology and attempted to see whether nanoparticle production can be upscaled.
    Anna Golova	Modeling experiments	Conducted several chemical and biological experiments to identify optimal temperature, pH and salt concentrations suitable for biological production of nanoparticles.
    Matthijs Hamstra	2022 iGEM InterLaboratory Study experiments and general laboratory assistance	Carried out the experiment for the InterLaboratoryStudy using the protocols and materials provided by iGEM and assisted with various experimental tasks in the lab.

    Fundraising and finance:

    Our team and finance manager, Mirthe Zandbelt, was responsible for actively obtaining funds for our project, managing project expenses, organizing meetings with potential sponsors, and working closely with various educational organizations to promote the project. Furthermore, she developed the crowdfunding page and successfully raised funds for the project.

    Design:

    The design manager, Moor de Waal, was in charge of designing the logo, animations, and styling the wiki. Throughout our project, Moor used Illustrator and After Effects to design flyers, posters and our promotion video. She worked closely with Maxim Velli who provided valuable insight for designing the wiki and promotion video.

    Collaborations:

    Our communication manager, Windi Putri Wulandari, was in charge of keeping track of any collaboration opportunities. With her assistance, our team was well informed about possible collaborations and often used these opportunities to work with different teams and to meet various iGEM teams from all around.

    Programming and Statistics:

    Matthijs Hamstra learned how to code during the iGEM project and successfully programmed our wiki. Nadine van de Brug and Anna Golova used the programming language R for data analysis.

    Outreach and Social Media:

    Our communication manager, Windi Putri Wulandari, was largely responsible for maintaining our social media platforms and keeping our followers updated. She used the platforms to promote science by working closely with Zainab Rashid, who designed science themed comics to spark an interest in younger students for synthetic biology and science in general. Windi also contacted media (newspapers and magazines) and student associations to promote our projects and crowdfunding. Furthermore, Mirthe Zandbelt worked with various organizations to promote our project through blogs and articles.

    Lab and safety:

    Tino Hoeksma was responsible for the safety in the laboratory, worked closely with our safety experts to ensure that the safety form was appropriately filled, and also ordered our lab supplies.

    Presentations:

    Our team presented at different symposiums or smaller meet-ups. All team members were involved in these presentations.

    Wiki:

    The wiki was programmed by Matthijs Hamstra and Maxim Velli and designed by Moor de Waal. All team members were involved in writing the text for the wiki, and the final text was mainly managed by Zainab Rashid and Maxim Velli.

    Wiki Pages:

    Page Name	Written by
    Attributions	Zainab Rashid
    Collaborations	Windi Putri Wulandari
    Contributions	Jennifer Adami, Mirthe Zandbelt, Anna Golova
    Crowdfunding	Mirthe Zandbelt
    Description	Zainab Rashid
    Engineering	Maxim Velli
    Entrepreneurship	Eva Pakvis
    Featured	Windi Putri Wulandari
    Implementation	Eva Pakvis
    Integrated Human Practices	Elise Stengs
    Interlab	Maxim Velli, Matthijs Hamstra
    Homepage	Moor de Waal
    Modeling	Anna Golova
    Notebook	Organized by Maxim Velli, Nadine van de Brug, Jennifer Adami
    Parts	Jennifer Adami
    Proof of concept	Nadine van de Brug, Moor de Waal
    Protocols	Jean Richard Quant, Jennifer Adami
    Public Engagement	Olivier Muzerie
    Results	Maxim Velli, Nadine van de Brug, Jennifer Adami, Zainab Rashid, Anna Golova
    Safety	Tino Hoeksma
    Sponsors	Mirthe Zandbelt
    Team Members	All

    Copyright attributions:

    • Icons: Flaticon
    • Icons: Freepik

    AOS