1. Biochemistry, structure, and cellular internalization of a four nanobody-bearing Fc dimer
- Author
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Estelle Marcheteau, Gilles Ferry, Charline Fagnen, Mathias Antoine, Eric Chabrol, Sophie Landron, Benjamin Fould, Jean A. Boutin, Catherine Vénien-Bryan, Johann Stojko, Sophie-Pénélope Guenin, Institut de Recherches SERVIER (IRS), Institut de minéralogie, de physique des matériaux et de cosmochimie (IMPMC), Muséum national d'Histoire naturelle (MNHN)-Institut de recherche pour le développement [IRD] : UR206-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université de Caen Normandie (UNICAEN), Normandie Université (NU), Roche Pharma Research and Early Development [Basel] (pRED), F. Hoffmann-La Roche [Basel], Institut de Recherches Internationales Servier [Suresnes] (IRIS), Pharmacochimie et Biologie pour le Développement (PHARMA-DEV), Institut de Recherche pour le Développement (IRD)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées
- Subjects
Receptor, ErbB-2 ,Dimer ,[SDV]Life Sciences [q-bio] ,Gene Expression ,Antigen-Antibody Complex ,Protein Engineering ,Biochemistry ,chemistry.chemical_compound ,subcellular localization ,characterization ,Cloning, Molecular ,Internalization ,media_common ,0303 health sciences ,biology ,Chemistry ,030302 biochemistry & molecular biology ,Multiple applications ,Recombinant Proteins ,Antibody ,Protein Binding ,Camelus ,medicine.drug_class ,media_common.quotation_subject ,Recombinant Fusion Proteins ,Full‐Length Papers ,Genetic Vectors ,VHH ,Monoclonal antibody ,03 medical and health sciences ,Antigen ,Cell Line, Tumor ,medicine ,Escherichia coli ,Animals ,Humans ,Amino Acid Sequence ,Antigens ,Molecular Biology ,030304 developmental biology ,cellular uptake ,Single-Domain Antibodies ,Trastuzumab ,Subcellular localization ,Immunoglobulin Fc Fragments ,Molecular Weight ,HER2 Antigen ,Biophysics ,biology.protein ,Protein Multimerization ,antibody like molecules - Abstract
International audience; VHH stands for the variable regions of heavy chain only of camelid IgGs. The VHH family forms a set of interesting proteins derived from antibodies that maintain their capacity to recognize the antigen, despite their relatively small molecular weight (in the 12,000 Da range). Continuing our exploration of the possibilities of those molecules, we chose to design alternative molecules with maintained antigen recognition, but enhanced capacity, by fusing four VHH with one Fc, the fragment crystallizable region of antibodies. In doing so, we aimed at having a molecule with superior quantitative antigen recognition (×4) while maintaining its size below the 110 kDa. In the present paper, we described the building of those molecules that we coined VHH2-Fc-VHH2. The structure of VHH2-Fc-VHH2 in complex with HER2 antigen was determined using electronic microscopy and modeling. The molecule is shown to bind four HER2 proteins at the end of its flexible arms. VHH2-Fc-VHH2 also shows an internalization capacity via HER2 receptor superior to the reference anti-HER2 monoclonal antibody, Herceptin®, and to a simple fusion of two VHH with one Fc (VHH2-Fc). This new type of molecules, VHH2-Fc-VHH2, could be an interesting addition to the therapeutic arsenal with multiple applications, from diagnostic to therapy.
- Published
- 2021
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