Fig. 5: Reliable in silico construction of scFv intrabodies.
From: Reliable repurposing of the antibody interactome inside the cell
a Design rules for reformatting an antibody as an optimised scFv intrabody. b Intracellular target specificity of 672 scFv constructed using the principles in (a). c Mean amino acid conservation for native to inverse folded scFv intrabody and charged residues in non-fixed positions within variable domain consensus sequences. d Charge frequency histograms for VL and VH framework sites before and after inverse folding. e Anti-p53 scFv 11D3 and 421 constructed using rules in (a) are highly soluble. Lower pane are sample processing controls. f Co-immunoprecipitation of p53 by anti-p53 scFv intrabodies. g Co-immunoprecipitation of UCHL1 by anti-UCHL1 scFv intrabodies. h Co-immunoprecipitation of GFP by anti-GFP scFv intrabody derived from Neuromabs antibody N86_38. All immunoprecipitation visualised by SDS-PAGE/western following immobilisation with anti-HA beads. Antibodies used to probe western membranes are stated to the left. IP Immunoprecipitation. Chothia numbering throughout. Western blots are representative of at least two experiments.
