Fig. 1

The production and identification of AAV2M^EC1 virus. A: In plasmids pEC1-VP2M and pR-VP1/3 M, the start codon of VP2 was mutated (labeled by asterisk) to prevent its expression. To eliminate binding to HSPG, R585 and R588 in pEC1-VP2M and pR-VP1/3 M were replaced with alanine (labeled by triangle). B: Schematic illustration of the AAV2M^EC1 capsid are shown. Five DARPin molecules (yellow) extending from the five-fold symmetry axis of the AAV2M^EC1 capsid, Illustration was created using Pymol and PDB files 6IH9 and 5KNG. C: Western blot analysis of AAV2M^EC1 virus. The fusion of DARPin molecule EC1 with VP2 protein led to a reduced mobility of AAV capsid proteins in SDS-PAGE. D: His-tag was inserted at the N-terminus of the EC1 sequence and packaged into virus, which was detected using His-tag antibody to detect EC1 protein in the virus. E: ELISA was used to determine EC1 display on the virus surface. AAV2M^{His − EC1} virus was able to bind to plates coated with His-tag antibody, while AAV2 virus was used as a control and the bound virus particles were quantified using Cap-specific antibody. N = 3 experiments; mean ± SD.