A novel, smaller scaffold for Affitins: Showcase with binders specific for EpCAM

Autor: Valentina Kalichuk, Véronique Préat, Gonzalo Obal, Frédéric Pecorari, Mike Maillasson, Federico Carrión, Barbara Mouratou, Ghislaine Béhar, Axelle Renodon-Cornière
Přispěvatelé: Nuclear Oncology (CRCINA-ÉQUIPE 13), Centre de Recherche en Cancérologie et Immunologie Nantes-Angers (CRCINA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Centre National de la Recherche Scientifique (CNRS)-Université d'Angers (UA), Advanced Drug Delivery and Biomaterials [Brussels, Belgium], Université Catholique de Louvain = Catholic University of Louvain (UCL)-Louvain Drug Research Institute (LDRI), Protein Biophysics [Montevideo] (UBP), Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), IMPACT - Interactions Moléculaires Puces ACTivités, European Erasmus Mundus Joint Doctorate in nanomedicine and pharmaceutical innovation, Nanofar, Bernardo, Elizabeth, Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes)-Université d'Angers (UA)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre hospitalier universitaire de Nantes (CHU Nantes)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Biotechnology and Bioengineering
Biotechnology and Bioengineering, Wiley, 2017, Epub ahead of print. ⟨10.1002/bit.26463⟩
Biotechnology and Bioengineering, 2017, Epub ahead of print. ⟨10.1002/bit.26463⟩
ISSN: 0006-3592
1097-0290
DOI: 10.1002/bit.26463⟩
Popis: International audience; Affitins are highly stable engineered affinity proteins, originally derived from Sac7d and Sso7d, two 7 kDa DNA-binding polypeptides from Sulfolobus genera. Their efficiency as reagents for intracellular targeting, enzyme inhibition, affinity purification, immunolocalization, and various other applications has been demonstrated. Recently, we have characterized the 7 kDa DNA-binding family, and Aho7c originating from Acidianus hospitalis was shown to be its smallest member with thermostability comparable to those of Sac7d and Sso7d. Here, after four rounds of selection by ribosome display against the human recombinant Epithelial Cell Adhesion Molecule (hrEpCAM), we obtained novel Aho7c-based Affitins. The binders were expressed in soluble form in Escherichia coli, displayed high stability (up to 74°C; pH 0-12) and were shown to be specific for the hrEpCAM extracellular domain with picomolar affinities (KD = 110 pM). Thus, we propose Aho7c as a good candidate for the creation of Affitins with a 10% smaller size than the Sac7d-based ones (60 vs. 66 amino acids).
Databáze: OpenAIRE
Externí odkaz: