Figure 8
From: Efficacy of an Adenoviral Vectored Multivalent Centralized Influenza Vaccine
Western Blot for Consensus HA Expression. 293 cells were infected with all 8 of the Ad vaccine vectors individually. The cells were harvested after 24 hrs. and cell lysates were run on a 8% SDS-PAGE gel. Protein was transferred to PVDF membranes and detected using polyclonal goat antiserum. Goats were immunized with A/PR/8/34, A/Singapore/1/57, A/Aichi/2/68, and A/Tern/S. Africa/61 for H1, H2, H3 and H5 protein expression, respectively. Due to differences in polyclonal anti-HA affinities the exposure times were different for each of the protein blots. A GAPDH control was also used as a standard. In order to determine strain-specific detection of HA proteins using polyclonal sera, the consensus proteins and wildtype virus proteins were probed with a combination of anti-H1, H3 and H5 goat polyclonal sera. The polyclonal antibody was detected by α-Goat-HRP secondary antibody. The blot was developed and imaged as one figure (B). The percent identity of the virus used to create the polyclonal sera relative to the antigen being detected is shown.
