Figure 1.

Approach for designing and sorting antibody libraries to identify antibodies with conformational and sequence specificity for Aβ fibrils. A parental antibody (scFv) was generated with Aβ residues 33–42 grafted into HCDR3 (red). Next, HCDR3 was diversified using natural diversity mutagenesis at 15 CDR sites, and the antibody library was displayed on the surface of yeast (S. cerevisiae). The library was then sorted using negative selections against disaggregated Aβ and positive selections against Aβ fibrils using antigens immobilized on magnetic beads. Finally, antibody variants from the enriched library were expressed as soluble proteins, and their conformational specificity was evaluated for Aβ fibrils relative to disaggregated Aβ.