FIGURE LEGENDS
Fig. 1: Target species study experimental design . US= ultrasound exam; EB= estradiol benzoate; EC= estradiol cypionate; P4= progesterone; IVD= intravaginal device.
Fig. 2: Culture of P5C3 clone in a 1 L bioreactor. Evolution of cell density, cell viability, lactate, glucose and reCG concentration along time. Temperature and perfusion rate per day are shown. Asterisk character indicates cell bleeding.
Fig. 3: SDS-PAGE of fractions corresponding to different stages of reCG purification. Samples were subjected to electrophoresis under non-reducing conditions and stained with Coomassie blue. Lane MW, Blue Plus® Protein Marker (Transgene Biotech); lane 1, clarified supernatant; lane 2, flowthrough; lane 3, wash; lane 4, eluate.
Fig. 4: Western blot analysis of reCG under different conditions. A: Unheated samples were electrophoresed under non-reducing conditions. B: Heated samples were electrophoresed under non‑reducing conditions. C: Heated samples were electrophoresed under reducing conditions. After SDS-PAGE proteins were probed with antiserum against reCG. For each panel, lane 1 corresponds to PMSG, lane 2 to reCG, lane 3 to supernatant of sCHOwt, and lane MW to ECLTM RainbowTM Marker (AmershamTM).
Fig. 5: IEF profile of PMSG (lane 1) and reCG (lane 2) glycoforms. After separation, proteins were transferred onto PVDF membrane and probed with rabbit polyclonal sera anti-reCG. The line indicates the range of pH employed.