Genetic Incorporation of Non-canonical Amino Acids in Anti-HER2 VHH: Expression and Characterization

Authors

  • Nuria Ramos Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France
  • Donna-Joe Bigot Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France
  • Gabrielle Zeder-Lutz Plateforme IMPReSs, UMR 7242, Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie de Strasbourg, 67412 Illkirch, France https://orcid.org/0000-0003-1959-1288
  • Jean-Marc Strub Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO), Institut Pluridisciplinaire Hubert Curien – IPHC. CNRS UMR 7178, Université de Strasbourg, France and Infrastructure Nationale de Protéomique ProFI, FR2048 CNRS CEA, 67037 Strasbourg, France https://orcid.org/0000-0001-6224-3428
  • Adilya Dagkesamanskaya Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France
  • Régis Fauré Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France https://orcid.org/0000-0002-5107-9009
  • Sébastien Nouaille Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France https://orcid.org/0000-0003-2038-098X
  • Cédric Y. Montanier Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France https://orcid.org/0000-0002-8205-7160
  • Gilles Ferry Pole d’expertise Biotechnologie, Chimie & Biologie, Institut de Recherches Servier, Croissy-sur-Seine, 78290 Croissy, France https://orcid.org/0000-0003-0689-7923
  • Renaud Wagner Plateforme IMPReSs, UMR 7242, Biotechnologie et Signalisation Cellulaire, Ecole Supérieure de Biotechnologie de Strasbourg, 67412 Illkirch, France https://orcid.org/0000-0002-4249-9853
  • Sarah Cianférani Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO), Institut Pluridisciplinaire Hubert Curien – IPHC. CNRS UMR 7178, Université de Strasbourg, France and Infrastructure Nationale de Protéomique ProFI, FR2048 CNRS CEA, 67037 Strasbourg, France https://orcid.org/0000-0003-4013-4129
  • Jean Albert Boutin Pole d’expertise Biotechnologie, Chimie & Biologie, Institut de Recherches Servier, Croissy-sur-Seine, 78290 Croissy, France https://orcid.org/0000-0003-0068-7204
  • Gilles Truan Toulouse Biotechnology Institute (TBI), Université de Toulouse, CNRS, INRAE, INSA, 31400 Toulouse, France https://orcid.org/0000-0002-3560-8302

DOI:

https://doi.org/10.37256/nat.5120243482

Keywords:

nanobodies, HER2, non-canonical amino acids, genetic incorporation, quality control, SPR

Abstract

Nanobodies– or VHH– are small proteins (~120 residues) issued from antibodies with an intact recognition for the original target of the antibody. In the present study, we show the possibility of incorporating non-canonical amino acids at precise location of the sequence via classical genetic techniques (Genetic Code Expansion). We demonstrate that the amount of recombinant protein obtained is compatible with large production format. We show that this protein can be purified, that its sequence corresponds to the theoretical molecular weight and that the two non-canonical amino acids are incorporated at the desired locations of the sequence. Finally, we show by SPR that the affinity of these VHHs is maintained towards its target, HER2.

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Published

2023-09-02

How to Cite

1.
Ramos N, Bigot D-J, Zeder-Lutz G, Strub J-M, Dagkesamanskaya A, Fauré R, Nouaille S, Montanier CY, Ferry G, Wagner R, Cianférani S, Boutin JA, Truan G. Genetic Incorporation of Non-canonical Amino Acids in Anti-HER2 VHH: Expression and Characterization. Nanoarchitectonics [Internet]. 2023 Sep. 2 [cited 2024 Apr. 18];5(1):24-42. Available from: https://ojs.wiserpub.com/index.php/NAT/article/view/3482