%0 Journal Article %T Impact of Site-Specific Conjugation of ScFv to Multifunctional Nanomedicines Using Second Generation Maleimide %+ Nanomédicaments et Nanosondes, EA 6295 (NMNS) %+ Groupe innovation et ciblage cellulaire (GICC), EA 7501 [2018-...] (GICC EA 7501) %+ Infectiologie et Santé Publique (UMR ISP) %A Hervé-Aubert, Katel %A Allard-Vannier, Emilie %A Joubert, Nicolas %A Lakhrif, Zineb %A Alric, Christophe %A Martin, Camille %A Viaud-Massuard, Marie-Claude %A Dimier-Poisson, Isabelle %A Aubrey, Nicolas %A Chourpa, Igor %Z "Region Centre-Val de Loire" (NCIS project) ; Ligue Nationale Contre le Cancer ; French Higher Education and Research ministry under the program "Investissements d'avenir" Grant Agreement: LabEx MAbImprove [ANR-10-LABX-53-01] and LabEx SynOrg [ANR-11-LABX-00-29] ; Tours University ; LabEx SynOrg %< avec comité de lecture %@ 1043-1802 %J Bioconjugate Chemistry %I American Chemical Society %V 29 %N 5 %P 1553-1559 %8 2018-03-23 %D 2018 %R 10.1021/acs.bioconjchem.8b00091 %M 29553717 %Z Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology %Z Life Sciences [q-bio]/Cancer %Z Life Sciences [q-bio]/Bioengineering/Biomaterials %Z Life Sciences [q-bio]/Pharmaceutical sciences/Medication %Z Chemical Sciences/Analytical chemistryJournal articles %X Biocompatible multifunctional nanomedicines (NMs) are known to be an attractive platform for targeted anticancer theranosis. However, these nanomedicines are of interest only if they efficiently target diseased cells and accumulate in tumors. Here we report the synthesis of a new generation of immunotargeted nanomedicines composed of a superparamagnetic iron oxide nanoparticle (SPION) core, polyethylene glycol coating and the anti-HER2 single chain fragment variable (scFv) of Trastuzumab antibody. We developed two novel bioengineered scFv carrying two cysteines located (i) at the end (4D5.1-cys2) or (ii) at the beginning (4D5.2-cys2) of its hexahistidine tag. The scFv bioconjugation was controlled via heterobifunctional linkers including a second generation maleimide (SGM). Our data indicated that the insertion of cysteines at the beginning of the hexahistidine tag was allowed to obtain nearly 2-fold conjugation efficiency (13 scFv/NP) compared to NMs using classical maleimide. As a result, the NMs-4D5.2 built using the optimal 4D5-cys2 and linkers equipped with SGM showed the enhanced recognition of HER2 in an ELISA format and on the surface of SK-BR-3 breast cancer cells in vitro. Their stability in serum was also significantly improved compared to the NMs-4D5. Our results showed the fundamental importance of the controlled ligand conjugation in the perspective of rational design of NMs with tailored physicochemical and biological properties. %G English %L hal-01757120 %U https://univ-tours.hal.science/hal-01757120 %~ UNIV-TOURS %~ INRA %~ AGREENIUM %~ INRAE %~ INFECTIOLOGIE-SANTE-PUBLIQUE %~ INRAEVALDELOIRE %~ TEST3-HALCNRS