Fig. 4

FPLC and Western blot analyses of partially purified recombinant un-tagged and tagged prorelaxin. (a) Human recombinant pro-relaxin H2 was purified from fermentation supernatant by ultrafiltration, IEX and RPC. The panel shows the RP chromatogram; the recovered fractions were analysed by western blotting. Lane 1) Unbound, lane 2) 20% B, lane 3) 30% B, lane 4) 35% B, M) molecular weight marker. (b) Human recombinant His tagged pro-relaxin H2 was recovered by affinity chromatography in batch and sequentially the sample was loaded on a RPC column; the resulting chromatogram is shown in the panel. The fractions recovered from RPC were analysed by western blotting. Lane 1) Unbound, lane 2) 20% B, Lane 3) 30% B, Lane 4) 35% B, M) molecular weight marker. The arrow in the RP chromatograms indicates the peaks recognized by the anti pro-relaxin antibody