Fig. 4. On-demand production of conjugate vaccines against F. tularensis using iVAX.

(A) iVAX reactions were prepared from lysates containing CjPglB and FtO-PS and primed with plasmid encoding immunostimulatory carriers, including those used in licensed vaccines. (B) We observed on-demand synthesis of anti-F. tularensis conjugate vaccines for all carrier proteins tested. Conjugates were purified using Ni-NTA agarose from 1-ml iVAX reactions lasting ~1 hour. Top panels show signal from probing with αHis to detect the carrier protein, middle panels show signal from probing with αFtO-PS, and bottom panels show αHis and αFtO-PS signals merged. Images are representative of at least three biological replicates. Dashed lines indicate that samples are from nonadjacent lanes of the same blot with the same exposure. Molecular weight ladders are shown at the left of each image.