The efficacy of DNA vaccination is enhanced in mice by targeting the encoded protein to dendritic cells
J. Clin. Invest. Godwin Nchinda, et al. 118:1427 doi:10.1172/JCI34224 [Go to this article.]

Figure 1
Design and characterization of antigen fused to single-chain antibody to DEC205. (A) Map of expression cassettes encoding vaccine protein (Vac protein) fused to single-chain scDEC and scControl. Heavy-chain (H) and light-chain (L) cDNAs of each mAb were connected by an interchain linker (ICL) and cloned in frame upstream of the cDNA for OVA, HIV gag fragments (p41, p24), or GFP in a eukaryotic expression plasmid containing the human CMV immediate early promoter and a bovine growth hormone polyadenylation signal (pAd). LP, leader peptide; Tag, myc and 6× histidine tag. (B) Binding to CHO cells expressing murine DEC205 (CHOmDEC205) or CHOneo control cells of recombinant single-chain mAbs-OVA fusion proteins detected by an OVA-specific, FITC-labeled secondary antibody. (C) Right part shows an overlay of the binding of the parental bivalent antibody (DEC-OVA) compared with scDEC-OVA. (D–F) Supernatants and lysates of 293T cells transfected with expression plasmids for scDEC or scControl conjugated to either OVA, HIV gag p41, HIV gag p24, or empty expression plasmid (pcDNA3.1; Invitrogen) were Western blotted using mAbs against OVA (D) or anti-HIV gag p24 (E and F).