Fiche publication
Date publication
août 2019
Journal
Protein science : a publication of the Protein Society
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CIANFERANI Sarah
,
Dr WAGNER Renaud
Tous les auteurs :
Hartmann L, Botzanowski T, Galibert M, Jullian M, Chabrol E, Zeder-Lutz G, Kugler V, Stojko J, Strub JM, Ferry G, Frankiewicz L, Puget K, Wagner R, Cianférani S, Boutin JA
Lien Pubmed
Résumé
In the continuous exploration of the VHH chemistry, biochemistry and therapeutic future use, we investigated two different production strategies of this small antibody-like protein, using an anti-HER2 VHH as a model. The total chemical synthesis of the 125 amino-acid peptide was performed with reasonable yield, even if optimization will be necessary to upgrade this kind of production. In parallel, we expressed the same sequence in two different hosts: E. coli and P. pastoris. Both productions were successful and led to a fair amount of VHHs. The integrity and conformation of the VHH were characterized by complementary mass spectrometry approaches, while surface plasmon resonance experiments were used to assess the VHH recognition capacity and affinity towards its "antigen". Using this combination of orthogonal techniques, it was possible to show that the 3 VHHs - whether synthetic or recombinant ones - were properly and similarly folded and recognized the "antigen" HER2 with similar affinities, in the nanomolar range. This opens a route towards further exploration of modified VHH with unnatural aminoacids and subsequently, VHH-drug conjugates. This article is protected by copyright. All rights reserved.
Mots clés
HER2, VHH, affinity, characterization, chemical synthesis, expression, ion-mobility, mass spectrometry, native chemical ligation, surface plasmon resonance
Référence
Protein Sci.. 2019 Aug 19;:
