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Next Generation Instrument for In-Depth Analysis of Mitochondrial Fitness

Periodic Reporting for period 2 - NextGen-O2k (Next Generation Instrument for In-Depth Analysis of Mitochondrial Fitness)

Reporting period: 2020-06-01 to 2021-10-31

Mitochondria and chloroplasts are key players when facing the new challenges of the millennium for biomedicine, biotechnology and for developing a sustainable bioeconomy. Monitoring mitochondrial fitness is important for diagnosis and therapeutic applications in many human pathologies, such as neurodegenerative diseases, obesity, diabetes, and cancer. In the biotechnology field, the optimization of algal growth rate and metabolite production requires novel investigations and monitoring of the bioenergetic pathway control exerted in respiration and photosynthesis. However, current technologies to study mitochondrial and chloroplast function are limited, providing segmented information, and lacking inter-laboratory reproducibility. As a result, the understanding of cellular bioenergetics for medical and algal biotechnology applications remains poor.

The overall objective of this project was to complete the development of the NextGen-O2k and prepare its market launch in 2021. The NextGen-O2k is a multi-sensor platform for in-depth studies of mitochondrial and chloroplast functional fitness. In a compact and affordable device, we have integrated several sensors for detecting multiple key parameters of bioenergetics, with superior resolution and reproducibility. With the NextGen-O2k we provide researchers with a tool able to improve knowledge about mitochondrial dysfunction and chloroplast photosynthesis versus dark respiration, while bringing savings in experimental time, samples and funds. The NextGen-O2k project was a step forward towards understanding mitochondrial implications in human pathologies and in the optimization of algal biomass production. Our focus is on generating knowledge about mitochondrial function and dysfunction, chloroplast photosynthesis and algal metabolism, while addressing the reproducibility crisis in science.
WP1: During the project, the new Q-, NADH and PB-Modules ― supported by the new architecture of the upgraded DatLab software ― have been developed and integrated. Additionally, we have successfully partnered with Key Opinion Leaders, whom we trained either on-site or virtually, for testing the Q- and PB-Modules. Feedback from the KOLs led to an additional photobiology light source (PBLS) and adjustment of the PB-Module. Furthermore, features of the new DatLab 8 software implemented by our subcontractor H-Tech were intensely tested and substrate-uncoupler-inhibitor-titration (SUIT) protocols were integrated.

WP2: Optimal measurement conditions were determined for using the Q-, NADH- and PB-Modules with a variety of biological samples. Simultaneously test programmes of the beta version of DatLab 8 including the new modules were conducted and improvements applied. New SUIT protocols were developed for general, pre-clinical, and algal biotechnology applications. A focus was also given to internal and external quality control measures, an important factor in the development of the NextGen-O2k instrument and relevant protocols.

WP3: In year one of the project, we aimed to improve the reproducibility of experimental assays and support customers with new developments. Therefore, we created an SOP for the new Q-Module, 6 new manuals and O2k-Videosupport clips for the Q-Module, DatLab7.4 mitochondrial preparations, and TPP+ and ion selective electrodes ISE. In the second year, we generated SOPs for the NextGen-O2k which were included in manuals and tutorial videos. These manuals and video clips, along with webinars, provide step-by step instructions, support, and information for using the NextGen-O2k.

WP4: As a cornerstone of our IP strategy, we successfully registered “NextGen-O2k”, “DatLab”, “High-Resolution Respirometry” and “O2k” as EU-trademarks. In addition to IP management and periodic IP Watch to ensure that our Freedom-to-operate (FTO) remains unimpeded, we increased our distributor engagement, helping us learning more about the market and customer needs. Furthermore, we explored the NextGen-O2k’s potential as a medical device and made progress on its algal biotechnology applications in collaboration with Key Opinion Leaders, which led to a publication. Although no certification for ISO is planned, several management documents have been created and implemented.

WP5: At the beginning of the project, we created a website to disseminate the results and also created a customer-friendly product website. For further strengthening our brand recognition, a communication strategy was put in place to bring an updated corporate identity with a new look to marketing materials and increased social media presence. Marketing materials, such as the NextGen-O2k folder and booth, were finalized and updated existing materials according to the latest corporate design. In addition to our print and digital presence, the Oroboros Team attended international conferences and hosted O2k-Workshops. Due to the COVID-19 pandemic, conference participation was restricted to virtual events and the concept of personalized individual training of the customers was implemented. KOLs, with whom we worked closely on prototype testing, were our first group to receive training in this format. To bring awareness to relevant scientific publications in the field, O2k-briefs summarizing key work were published regularly. Furthermore, an open access peer-reviewed journal, Bioenergetics Communications, was initiated. linked to the MitoFit Preprints server. Several scientific and methodological papers were published by our group or in collaboration with KOLs.
The all-in-one concept of the NextGen-O2k offers a unique instrument for simultaneous measurements of O2 flux and further parameters. As a cost-efficient, powerful, and compact equipment for space-constrained laboratories, it overcomes existing market restraints. In the biomedical field, the NextGen-O2k will revolutionize the current research methods, providing deeper insight into cellular functions in health and disease with the Q- and NADH-modules.

The instrumentation market for measurement of photosynthesis and bioenergetics in microalgae and plant cells is enriched by the PB-Module. With the support of the Key Opinion Leaders, several protocols were developed to facilitate the entry into this market and results published in journals. Moreover, our updated IPR strategy and solid distributor network including Asia, South America and Middle East increase the market reach of the NextGen-O2k.

The NextGen-O2k was introduced to the scientific community at the NextGen-O2k Summit. The integrated modules (Q for measuring Q-redox states, NADH for NAD(P)H redox states and PB for photosynthesis) are ready to be applied in research institutions and industry in the biomedical and algal biotechnological fields.

With the availability of the NextGen-O2k, the quality and reproducibility of respirometric measurements in both fields will be significantly improved. As a result, the current knowledge of cell physiology will be improved and factors affecting algal biomass production, helping push forward the European bioeconomy based on algae-derived products and positively impacting the society with the development of a greener economy. The NextGen-O2k will empower laboratories all over the world to contribute to solving the societal challenges of the 21st century
Distributor engagement Analytics China 2020
Training on Q-Module
Q-Module training of KOLs
ASMRM2019 meeting with distributor Berthold Japan K.K.
NextGen-O2k with Photobiology-Module in Oroboros O2k-Laboratory
Oroboros exhibition booth
Virtual workshop
NextGen-O2k awarded as best Tyrolean innovation in Life-Sciences 2019
PB-Module measurements
Behind the scenes: NextGen-O2k Summit