Fig. 1: Nanobody selection against α-syn PFF.
From: α-Synuclein fibril-specific nanobody reduces prion-like α-synuclein spreading in mice
a Nanobody crystal structure (based on PDB: 4LDE). Conserved disulfide bond (orange) was removed in our libraries. b Nanobody selection schematics. Nanobodies were expressed on the yeast surface by fusion to the C-terminus of Aga2p protein, followed by the FLAG tag. Yeast cells that bind to α-syn PFF were selected using MACS and FACS. Red trapezoid, a representative selection gate. c FACS analysis of nanobody libraries before and after selection. After 6 rounds of selection, the selected nanobodies showed a higher PFF-binding signal, compared to the original libraries and negative control (without α-syn PFF). The numbers in the upper right Q2 indicate the yeast population ratio of Q2/Q4.
