Site-directed mutagenesis for improving biophysical properties of VH domains

Download
  1. Get@NRC: Site-directed mutagenesis for improving biophysical properties of VH domains (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1007/978-1-60761-652-8_22
AuthorSearch for: ; Search for: ; Search for:
TypeArticle
Journal titleMethods in Molecular Biology
Volume634
Issue5
Pages309330; # of pages: 22
SubjectIn vitro mutagenesis; Splice overlap extension; VH; Biophysical properties; Reversible thermal unfolding; Nonaggregating; Surface plasmon resonance; Circular dichroism
AbstractRecombinant antibody fragments are significant therapeutic and diagnostic reagents. As such, their efficacy depends heavily on their affinities and biophysical properties. Thus, mutagenesis approaches have been extensively applied to recombinant antibodies to improve their affinity, stability, and solubility. Among the existing recombinant antibody variants, human VH domains stand out as the ones with the general need of solubility engineering at some point during their development; this solubility engineering step transforms VHs into nonaggregating, functional entities, rendering them useful as therapeutic and diagnostic reagents. Here, we present one of several approaches that have been employed to develop nonaggregating human VH domains. We apply an in vitro site-directed mutagenesis approach to an aggregating human VH domain by means of a splice overlap extension technique. The resultant mutant VHs are nonaggregating in contrast to the parent wild type VH and less prone to aggregation following thermal unfolding.
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedYes
NPARC number17401053
Export citationExport as RIS
Report a correctionReport a correction
Record identifiera32261d2-c2fd-4090-ab61-d1ebe1a813f2
Record created2011-03-25
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)