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Figure 8: FSH characterization by size exclusion chromatography and Western blot analysis. Samples of hFSH24, hFSH24/21, and hFSH21 were applied to a 1 x 30 cm Superdex 75 column and the chromatogram developed with 20% acetonitrile in 0.2 M ammonium bicarbonate at a flow rate of 0.4 ml/min. Retention times (min) for each FSH peak are indicated. A. Fully-glycosylated hFSH24 from Superdex 75 fraction A (Figure 1G). Inset: Lane 1, fraction A; lane 2, fraction A’; lane 3, fraction B; lane 4, fraction B’; lane 5, fraction C; lane 6, fraction C’; lane 7, purified pituitary hFSH24/21. B. Mixture of both hFSH glycoforms. C. Hypo-glycosylated hFSH21 from fraction C, Figure G. |