000095476 001__ 95476
000095476 005__ 20230622083311.0
000095476 0247_ $$2doi$$a10.3390/ijms21176165
000095476 0248_ $$2sideral$$a119532
000095476 037__ $$aART-2020-119532
000095476 041__ $$aeng
000095476 100__ $$0(orcid)0000-0002-1657-4792$$aGuerrero-Ochoa, Patricia$$uUniversidad de Zaragoza
000095476 245__ $$aProduction of a granulysin-based, tn-targeted cytolytic immunotoxin using pulsed electric field technology
000095476 260__ $$c2020
000095476 5060_ $$aAccess copy available to the general public$$fUnrestricted
000095476 5203_ $$aGranulysin 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.
000095476 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/SAF2016-76338-R$$9info:eu-repo/grantAgreement/ES/MINECO/PID2019-105451GB-I00$$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2013-44367-C2-2-P$$9info:eu-repo/grantAgreement/ES/MINECO/BFU2016-75633-P$$9info:eu-repo/grantAgreement/ES/DGA/E34-R17$$9info:eu-repo/grantAgreement/ES/DGA/B31-17R
000095476 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000095476 590__ $$a5.923$$b2020
000095476 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b67 / 297 = 0.226$$c2020$$dQ1$$eT1
000095476 591__ $$aCHEMISTRY, MULTIDISCIPLINARY$$b49 / 178 = 0.275$$c2020$$dQ2$$eT1
000095476 592__ $$a1.455$$b2020
000095476 593__ $$aCatalysis$$c2020$$dQ1
000095476 593__ $$aComputer Science Applications$$c2020$$dQ1
000095476 593__ $$aInorganic Chemistry$$c2020$$dQ1
000095476 593__ $$aSpectroscopy$$c2020$$dQ1
000095476 593__ $$aMolecular Biology$$c2020$$dQ1
000095476 593__ $$aOrganic Chemistry$$c2020$$dQ1
000095476 593__ $$aPhysical and Theoretical Chemistry$$c2020$$dQ1
000095476 593__ $$aMedicine (miscellaneous)$$c2020$$dQ1
000095476 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000095476 700__ $$aAguilar-Machado, D.
000095476 700__ $$aIbáñez-Pérez, Raquel$$uUniversidad de Zaragoza
000095476 700__ $$0(orcid)0000-0001-6815-6720$$aMacías-León, Javier$$uUniversidad de Zaragoza
000095476 700__ $$0(orcid)0000-0002-3122-9401$$aHurtado-Guerrero, Ramón$$uUniversidad de Zaragoza
000095476 700__ $$0(orcid)0000-0003-3957-9091$$aRaso, Javier$$uUniversidad de Zaragoza
000095476 700__ $$0(orcid)0000-0002-5175-8394$$aAnel, Alberto$$uUniversidad de Zaragoza
000095476 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000095476 7102_ $$11002$$2050$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Biología Celular
000095476 7102_ $$12008$$2780$$aUniversidad de Zaragoza$$bDpto. Produc.Animal Cienc.Ali.$$cÁrea Tecnología de Alimentos
000095476 773__ $$g21, 17 (2020), 6165 1-13$$pInt. j. mol. sci.$$tInternational Journal of Molecular Sciences$$x1661-6596
000095476 8564_ $$s1995302$$uhttps://zaguan.unizar.es/record/95476/files/texto_completo.pdf$$yVersión publicada
000095476 8564_ $$s482277$$uhttps://zaguan.unizar.es/record/95476/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000095476 909CO $$ooai:zaguan.unizar.es:95476$$particulos$$pdriver
000095476 951__ $$a2023-06-21-14:59:53
000095476 980__ $$aARTICLE