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After 45 h, the conjugates were isolated by semi-preparative HPLC from the crude reaction mixtures with using a Zorbax Eclipse XDB-C18 HPLC column (80 , 9.4 250 mm, 5 m) at a flow rate of 4 mL/min with acetonitrile (solvent A, containing 0.1% v/v TFA) and Milli Q water (solvent B, containing 0.1% v/v TFA)