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Isolation of monomeric human VHS by a phage selection

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1074/jbc.M509900200
AuteurRechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 2; Rechercher : 2; Rechercher : 2; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut des sciences biologiques du CNRC
  2. Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
FormatTexte, Article
Sujet61-GH3P6-1-1-1-1-1-G-E; genome; pharmaceutical; protein; proteins; isolation; selection
Résumé
Human V(H) domains are promising molecules in applications involving antibodies, in particular, immunotherapy because of their human origin. However, they are, in general, prone to aggregation. Therefore, various strategies have been employed to acquire monomeric human V(H)s. We had previously discovered that filamentous phages displaying engineered monomeric V(H) domains gave rise to significantly larger plaques on bacterial lawns than phages displaying wild type V(H)s with aggregation tendencies. Using plaque size as the selection criterion and a phage-displayed naive human V(H) library we identified 15 V(H)s that were monomeric. Additionally, the V(H)s demonstrated good expression yields, good refolding properties following thermal denaturation, resistance to aggregation during long incubation at 37 °C, and to trypsin at 37 °C. These 15 V(H)s should serve as good scaffolds for developing immunotherapeutics, and the selection method employed here should have general utility for isolating proteins with desirable biophysical properties.
Date de publication
Maison d’éditionAmerican Society for Biochemistry and Molecular Biology
Dans
  • The Journal of Biological Chemistry 280, nº 50 : 41395–41403.
Langueanglais
Publications évaluées par des pairsOui
Numéro du CNRCTO2005
NRC-BRI-42506
Numéro NPARC3539892
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Identificateur de l’enregistrement9ed11f6a-a5ec-476d-950e-0490b7220833
Enregistrement créé2009-03-01
Enregistrement modifié2020-04-07
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