Isolation of monomeric human VHS by a phage selection

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DOIResolve DOI: http://doi.org/10.1074/jbc.M509900200
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TypeArticle
Journal titleThe Journal of Biological Chemistry
Volume280
Issue50
Pages4139541403; # of pages: 9
Subject61-GH3P6-1-1-1-1-1-G-E; genome; human; pha; protein; proteins; isolation; selection
AbstractHuman 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
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada; NRC Institute for Biological Sciences
Peer reviewedNo
NRC numberTO2005
42506
NPARC number3539892
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Record identifier9ed11f6a-a5ec-476d-950e-0490b7220833
Record created2009-03-01
Record modified2016-05-09
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