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. 2011 Nov 11;6(11):e27666. doi: 10.1371/journal.pone.0027666

Figure 1. Recombinants produced using E. coli cell-free IVTT system.

Figure 1

Western Blot and quantification of (A) viral antigens FluM, FluHA, CMVpp65 or EBNA3A and (B) parasite antigen PyCSP pIVEX HisHA IVTT products, probed with mAb against the C-terminal HA tag. Whole IVTT extracts (5 µl) of each antigen were run on a 12% NUPAGE gel, transferred to a PVDF membrane, and probed with anti-HA HRP antibody (1∶500 dilution). Protein expression was quantitated against an IVTT-produced recombinant P. falciparum (PF14_0051) protein of known concentration expressing the same N-terminal 6xHis and C-terminal HA tags. Yields for each full length antigen were as follow: FluM, 1.15 µg/ µl; FluHA, 0.73 µg/ µl; CMVpp65, 0.7 µg/ µl; EBNA3A, 4 ng/ µl; and PyCSP, 0.6 µg/ µl.