Fig. 1
ScFv-MC1 design, expression and characterization. a scFv-MC1 diagram: 5’-terminal signal peptide (SP), variable heavy (VH) and variable light (VL) chains joined together by a 15 amino acid residue linker (Gly4Ser)3, and 3’-terminal Myc and His6X tags. b HEK293 cells were transiently transfected with empty vector (empty v.) or scFv-MC1. In the cell lysate (immunoblotting) the scFv-MC1 was detected as a 30 kDa band; c in the conditioned medium the presence of scFv-MC1 was assessed using a specific ELISA and was expressed as ODs (optical densities). Cells transfected with the empty vector did not show any sign of reactivity in western blotting or ELISA. d Purified scFv-MC1, run on SDS-PAGE gel and stained with Coomassie: two different purified preparations (1, 2) revealed the expected molecular weight (30 kDa). e scFv-MC1 antigen-binding specificity: purified scFv-MC1 tested for its activity on the specific peptide (998b) was compared to the MC1 mAb, showing a comparable reactivity; CP13 and DA31 were used as negative controls. f Representative images of AD brain immunohystochemistry: both scFv-MC1 and MC1 (1:500 dilution) showed specificity for the neurofibrillary pathology (Olympus BH-2 microscope, bar: 50 μm)