ScFv antibody fragments are a promising alternatives to full-length antibodies for both therapeutic and diagnosis applications. They can be overexpressed in bacteria, which enables easy large scale production. Since scFv are artificial constructs, they are poorly soluble and prone to aggregation, which makes them difficult to manipulate and to refold. Here, we report stabilization and refolding of scFv fragments from urea-unfolded solutions based on the use of micromolar amounts of polymers playing the role of artificial chaperons. Using fluorescence correlation spectroscopy, we determined the size and aggregation number of complexes of scFv with unmodified or hydrophobically-modified poly(sodium acrylate). The evolution of the secondary structure along the refolding procedure, in the presence or absence of 0.4 M L-arginine at scFv:polymer < 1:5 wt/wt, was determined by high-sensitivity synchrotron-radiation circular dichroism, SRCD. Measurements revealed that refolding in the presence of polymers yields native-like secondary structure, though a different folding pathway can be followed compared to refolding in the absence of polymer. This is the first report on the use of macromolecular additives to assist refolding of a multi-domain protein of therapeutic interest.3
Back Cover: Aggregation‐prone antibody single‐chain variable fragments (scFv) are refolded from urea solutions in the presence of amphiphilic derivatives of poly(sodium acrylate) playing the role of artificial chaperons. Weak noncovalent interactions with the polymers impart solubility to partly‐folded scFv intermediates, resulting in recovery of the native‐like structure. This is reported by Nicolas Martin, Narciso Costa, Frank Wien, Françoise M. Winnik, Céline Ortega, Amaury Herbet, Didier Boquet, and Christophe Tribet, article number 1600213.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.