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Isolation of monomeric human VHS by a phage selection

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1074/jbc.M509900200
AuthorSearch for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 2; Search for: 2; Search for: 2; Search for: 1
Affiliation
  1. National Research Council of Canada. NRC Institute for Biological Sciences
  2. National Research Council of Canada. NRC Biotechnology Research Institute
FormatText, Article
Subject61-GH3P6-1-1-1-1-1-G-E; genome; pharmaceutical; protein; proteins; isolation; selection
Abstract
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.
Publication date
PublisherAmerican Society for Biochemistry and Molecular Biology
In
  • The Journal of Biological Chemistry 280, no. 50: 41395–41403.
LanguageEnglish
Peer reviewedYes
NRC numberTO2005
NRC-BRI-42506
NPARC number3539892
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Record identifier9ed11f6a-a5ec-476d-950e-0490b7220833
Record created2009-03-01
Record modified2020-04-07
Date modified: