Percorrer por autor "Komporday, Katalin Luca"
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- Development of IL-11 receptor targeting miniproteinsPublication . Komporday, Katalin Luca; Oroszlan, Gabor; Power, Deborah MaryDe novo protein design has emerged as a transformative approach for creating novel therapeutic molecules with highly specific functions and tailored structures. This thesis presents the design and development of miniproteins targeting the interleukin-11 receptor alpha (IL-11Rα), a key mediator in fibrotic diseases such as idiopathic pulmonary fibrosis (IPF). Miniproteins, small stable peptides with well-defined tertiary structures, offer advantages over traditional therapies by small molecules or antibodies through higher selectivity, improved tissue penetration, and lower risk of immunogenicity. Leveraging AI-driven bioinformatics tools, a library of 400 different miniprotein scaffolds was designed to selectively bind the IL-11Rα, competing with its natural ligand, IL-11 cytokine. The miniprotein library was synthesized as an oligonucleotide pool, cloned into phagemid vectors, and expressed on the pIII M13 bacteriophage surface for phage display-based selection. A biopanning campaign was used to enrich miniproteins with strong binding affinities for IL-11Rα across multiple screening rounds. Two lead candidates, DN226 and DN213 demonstrated high specificity and affinity confirmed by ELISA assays. However, DN226 showed a better affinity and competitiveness against the IL-11 cytokine for binding to the receptor compared to DN213. Since transient transfection expression of IL-11Rα in FreeStyle Human Embryonic Kidney (HEK293-F) cells faced yield and aggregation challenges, commercially sourced IL-11Rα was employed in downstream assays. The combination of computational design with classical biotechnological approaches enabled the rapid identification of functional miniproteins capable of binding the IL-11Rα. This work underscores the potential of AI-driven de novo protein design as a versatile technology for precise drug discovery and development. The identified miniproteins provide promising hit molecules for developing innovative, targeted anti-fibrotic therapies. Future work should focus on optimizing miniprotein candidates and evaluating their efficacy in cell-based and in vivo studies, paving the way for novel treatment approaches for fibrotic diseases, like IPF.