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Processing of proHD5 by trypsin at 30 min and 2 h. HPLC analysis was performed at 40 °C on a Waters XBridge C18 column (3.5 μm, 4.6 × 150 mm) using a linear gradient of 5–45% acetonitrile containing 0.1% trifluoroacetic acid at a flow rate of 1 ml/min over 30 min. The chromatographic peaks were identified by ESI-MS analyses aided by simulated trypsin digestion. mAU, milliabsorbance units.
