Human apolipoprotein E (ApoE) exists as three common isoforms that differ by single amino acid substitutions. ApoE4 (Arg112/Arg158) is the strongest genetic risk factor for late-onset Alzheimer's disease (LOAD)-a significant cause of mortality and disability worldwide. At the protein level, previous studies suggest that the conformational landscape of ApoE4 is different to that of the wild-type ApoE3 (Cys112/Arg158). To date, however, effective ligands capable of selectively targeting the ''ApoE4-conformation'' remain elusive. In the present study we sought to use phage display technology to isolate several single domain antibody fragments (i.e. V NAR ), initially derived from the wobbegong shark, that are reactive against human ApoE isoforms. To this end we have expressed, purified, and characterized recombinant ApoE3 and ApoE4 for use as bait in phage display. After 3-5 rounds of panning enrichment against ApoE isoforms was observed. From these rounds of panning several unique clones were identified and subsequently subcloned, expressed, and purified to near homogeneity. We are currently in the process of more fully characterising the binding propensities of these clones.
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.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.