2026-04-14T19:29:52Zhttps://uvadoc.uva.es/oai/request

oai:uvadoc.uva.es:10324/741942025-05-09T10:48:46Zcom_10324_1134com_10324_931com_10324_894col_10324_1213

Tavladoraki, Paraskevi Girotti ., Alessandra Donini, Marcello Arias Vallejo, Francisco Javier Mancini, Camillo Veronica Morea Roberta Chiaraluce Valerio Consalvi Eugenio Benvenuto 2025-01-21T16:01:41Z 2025-01-21T16:01:41Z 1999 European Journal of Biochemistry (FEBS Journal). 262, 617-624 0014-2956 https://uvadoc.uva.es/handle/10324/74194 10.1046/j.1432-1327.1999.00443.x 617 262 624 FEBs Journal European Journal of Biochemistry 2 Producción Científica Despite the well-known crucial role of intradomain disulfide bridges for immunoglobulin folding and stability, thesingle-chain variable fragment of the anti-viral antibody F8 is functionally expressed when targeted to the reducingenvironment of the plant cytoplasm. We show here that this antibody fragment is also functionally expressed in thecytoplasm of Escherichia coli. A gel shift assay revealed that the single-chain variable fragment (scFv) accumulatingin the plant and bacterial cytoplasm bears free sulfhydryl groups. Guanidinium chloride denaturation/renaturationstudies indicated that refolding occurs even in a reducing environment, producing a functional molecule with thesame spectral properties of the native scFv(F8). Taken together, these results suggest that folding and functionality ofthis antibody fragment are not prevented in a reducing environment. This antibody fragment could thereforerepresent a suitable framework for engineering recombinant antibodies to be targeted to the cytoplasm. (FAIR1-CT95-0905) from the European Commission (to E.B.) and the Italian Ministero dell’Università e della Ricerca Scientifica e Tecnologica (MURST). European Commission. application/pdf eng Wiley info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ Attribution-NonCommercial-NoDerivatives 4.0 Internacional Intrabody, scFv, (ScFv anticuerpos intracelulares, estabilidad proteinas A single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion SI