https://phaidra.vetmeduni.ac.at/o:4686 Encapsidic production and isolation of degradation-prone polypeptides Gawin, Agnieszka Agnieszka Gawin 0000-0002-1457-6049 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Pankowski, Jedrzej Jedrzej Pankowski 0000-0003-3968-1790 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Zarechyntsava, Maria Maria Zarechyntsava https://orcid.org/0009-0006-3864-4559 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Kwasna, Dominika Dominika Kwasna https://orcid.org/0000-0002-6792-7703 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Kloska, Damian Damian Kloska https://orcid.org/0000-0003-3206-5493 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Koziej, Lukasz Lukasz Koziej https://orcid.org/0000-0003-3885-3845 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Glatt, Sebastian Sebastian Glatt https://orcid.org/0000-0003-2815-7133 Department for Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria Kachamakova-Trojanowska, Neli Neli Kachamakova-Trojanowska https://orcid.org/0000-0002-3226-0726 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Azuma, Yusuke Yusuke Azuma https://orcid.org/0000-0003-3543-3159 Malopolska Centre of Biotechnology, Jagiellonian University, Krakow 30-387, Poland Royal Society of Chemistry (RSC) journal article open access eng Degradation during production and delivery is a significant bottleneck in developing biomolecular therapies. Protein cages, formed by engineered variants of lumazine synthase, present an effective strategy for the microbial production and isolation of labile biomolecular therapies. Genetic fusion of the target polypeptide to a cage component protomer ensures its efficient encapsulation within the cage during production in host bacterial cells, thereby protecting it from degradation. Furthermore, controlled cage opening outside the cellular environment facilitates the isolation of the encapsulated cargo through sequence-specific protease cleavage. Notably, the system features a modular patchwork assembly to prevent guest overloading, avoiding unwanted incomplete cage formation and insoluble aggregates. The broad applicability of the “encapsidic” approach is demonstrated by the efficient production of six distinct, intrinsically disordered polypeptides with proven therapeutic potential. http://creativecommons.org/licenses/by/3.0/ https://phaidra.vetmeduni.ac.at/api/object/o:4686/download 10.1039/d5tb01033k https://phaidra.vetmeduni.ac.at/o:605 application/pdf Journal of Materials Chemistry B issn:2050-750X VoR Journal of Materials Chemistry B 13 39 12605 12613 2.25 MB 2025