Periodic Reporting for period 1 - INCYPROnext (INCYPRO: A key technology to enable the broad application of proteins in diagnostics and therapeutics)
Berichtszeitraum: 2022-06-01 bis 2023-05-31
Proteins are not only one of life’s main building blocks, but also key tools for many industrial applications, ranging from drug discovery and diagnostics to chemical production processes. The main limitation of proteins is their low stability at non-physiological conditions, as they are commonly required in industrial, medical and biotechnological processes. Consequently, proteins often loose their structural integrity and thus their activity under application-relevant conditions. There are thousands of proteins with large potential, for instance in the context of protein targets related to diseases with high unmet need, point-of-care diagnostics and therapeutic proteins. However, many cannot be utilized due to a lack of generally applicable technologies that allow straight-forward and meaningful protein stabilization. To address the challenges associated with protein stabilization, a versatile technology overcoming costly, time-consuming and uncertain trial-and-error approaches is required. The INCYPROnext project aims to provide a universal solution to the stabilization of proteins for biotechnological and biomedical applications. In the frame of the ERC PoC project, INCYPRO, we have developed a radically new technology for stabilizing proteins that has been validated in lab. Our innovation combines structural and molecular biology as well as chemistry to provide a highly effective and efficient technology for the stabilization of a broad range of proteins that are currently impossible to stabilize, delivering minimally modified proteins with a highly stabilized structure and retained functionality. This is an ideal value proposition to play a key role in a growing market, which can allow us to both offer B2B services to customers in the pharmaceutical, biotech, research and industrial processes segments, as well as to start our own internal drug discovery and development pipeline. In this EIC Transition project, we will validate the INCYPRO technology on a range of different proteins in a real operating environment (TRL5).
For the broad application of the INCYPRO technology, we are establishing a time-efficient workflow comprised of several steps that require optimization and standardization.We have started with the establishment of an accurate computational tool for the design and prediction of sites for INCYPRO modification. The resulting protein variants then go into our production process. Here, the workflow for protein expression, INCYPRO crosslinking and biophysical characterisations have been established and standardized to accommodate processing of several protein variants in parallel. Selected INCYPRO-stabilized proteins for the development of therapeutic proteins will be tested towards efficacy, toxicity and immunogenicity. Further in the project, the upscaling of crosslinker synthesis and production of selected INCYPRO-stabilized proteins is planned.
We have successfully stabilized various proteins with the INCYPRO technology and are continuing to validate the technology on a broad range of different protein structures. We show that we can add stability to proteins that had already been engineered by mutation. We show that INCYPRO can enable enzymes to be active under normally denaturing conditions. We were able to shift the melting temperature of a protein by +39°C, and we show that an INCYPRO-stabilized enzyme stays active 20 days longer at elevated temperatures than its unmodified counterpart. Together, these findings underline that our structural stabilization approach efficiently translates into enhanced activity. While we will continue our approach to stabilize different protein structures and use the generated data to improve our computational design and prediction module, we will start building a development pipeline. In strategic collaborations with partners, selected proteins will be developed towards therapeutic protein candidates, stabilized target proteins for drug development, and diagnostic tools.