95476 20230622083311.0 doi 10.3390/ijms21176165 sideral 119532 ART-2020-119532 eng Guerrero-Ochoa, Patricia Universidad de Zaragoza (orcid)0000-0002-1657-4792 Production of a granulysin-based, tn-targeted cytolytic immunotoxin using pulsed electric field technology 2020 Access copy available to the general public Unrestricted Granulysin is a protein present in the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, with cytolytic activity against microbes and tumors. Previous work demonstrated the therapeutic effect of the intratumoral injection of recombinant granulysin and of the systemic injection of an immunotoxin between granulysin and the anti-carcinoembryonic antigen single-chain Fv antibody fragment MFE23, which were produced in the yeast Pichia pastoris. In the present work, we developed a second immunotoxin combining granulysin and the anti-Tn antigen single-chain Fv antibody fragment SM3, that could have a broader application in tumor treatment than our previous immunotoxin. In addition, we optimized a method based on electroporation by pulsed electric field (PEF) to extract the remaining intracellular protein from yeast, augmenting the production and purificiation yield. The immunotoxin specifically recognized the Tn antigen on the cell surface. We also compared the thermal stability and the cytotoxic potential of the extracellular and intracellular immunotoxins on Tn-expressing human cell lines, showing that they were similar. Moreover, the bioactivity of both immunotoxins against several Tn+ cell lines was higher than that of granulysin alone. info:eu-repo/grantAgreement/ES/DGA/B31-17R info:eu-repo/grantAgreement/ES/DGA/E34-R17 info:eu-repo/grantAgreement/ES/MINECO/BFU2016-75633-P info:eu-repo/grantAgreement/ES/MINECO/CTQ2013-44367-C2-2-P info:eu-repo/grantAgreement/ES/MINECO/PID2019-105451GB-I00 info:eu-repo/grantAgreement/ES/MINECO/SAF2016-76338-R info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.923 2020 BIOCHEMISTRY & MOLECULAR BIOLOGY 67 / 297 = 0.226 2020 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 49 / 178 = 0.275 2020 Q2 T1 1.455 2020 Catalysis 2020 Q1 Computer Science Applications 2020 Q1 Inorganic Chemistry 2020 Q1 Spectroscopy 2020 Q1 Molecular Biology 2020 Q1 Organic Chemistry 2020 Q1 Physical and Theoretical Chemistry 2020 Q1 Medicine (miscellaneous) 2020 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Aguilar-Machado, D. Ibáñez-Pérez, Raquel Universidad de Zaragoza Macías-León, Javier Universidad de Zaragoza (orcid)0000-0001-6815-6720 Hurtado-Guerrero, Ramón Universidad de Zaragoza (orcid)0000-0002-3122-9401 Raso, Javier Universidad de Zaragoza (orcid)0000-0003-3957-9091 Anel, Alberto Universidad de Zaragoza (orcid)0000-0002-5175-8394 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 1002 050 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Biología Celular 2008 780 Universidad de Zaragoza Dpto. Produc.Animal Cienc.Ali. Área Tecnología de Alimentos 21, 17 (2020), 6165 1-13 Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 1995302 http://zaguan.unizar.es/record/95476/files/texto_completo.pdf Versión publicada 482277 http://zaguan.unizar.es/record/95476/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:95476 articulos driver 2023-06-21-14:59:53 ARTICLE