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Updated: Jun 10, 2026

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
Published on: January 26, 2017
Site-directed mutagenesis for improving biophysical properties of VH domains
Mehdi Arbabi-Ghahroudi1, Roger Mackenzie, Jamshid Tanha
1Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON, Canada.
Abstract:
Recombinant 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 V(H) domains stand out as the ones with the general need of solubility engineering at some point during their development; this solubility engineering step transforms V(H)s 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 V(H) domains. We apply an in vitro site-directed mutagenesis approach to an aggregating human V(H) domain by means of a splice overlap extension technique. The resultant mutant V(H)s are nonaggregating in contrast to the parent wild type V(H) and less prone to aggregation following thermal unfolding.
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