Fig. 1.

Design, expression, and purification of an anti-vGAT Fab domain (a) Expression plasmids encoding for the Fab heavy and light chains. Heavy chain and light chains contained a BM40 signal sequence which facilitated secretion of expressed protein into the cell culture media for easy purification. Both constructs included a variable region and constant region, and the heavy chain additionally included a CDK (Cysteine-Glutamate-Lysine) motif for Fab dimerization, and for affinity purification, a hexahistidine (6-HIS) tag followed by a Twin-Strep tag. Cartoon schematic (BioRender) of the engineered recombinant anti-vGAT Fab domain dimer used for isolation of native vGAT synaptic vesicles. (b) Coomassie-stained SDS PAGE analysis of the purified anti-vGAT Fab region.