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The resin was washed by 20 column volumes of wash buffer A [20 mM HEPES pH 7.5, 500 mM NaCl, 2% (w/v) glycerol, 0.05% (w/v) DDM, 0.01% (w/v) CHS and 30 mM imidazole] and 10 column volumes of wash buffer B [20 mM HEPES, pH 7.5, 500 mM NaCl, 2% (w/v) glycerol, 0.02% (w/v) DDM, 0.01% (w/v) CHS and 50 mM imidazole], followed by incubation with Fab7F38 at a molar ratio of 1: 1.5 in 3 mL wash buffer C [20 mM HEPES pH 7.5, 500 mM NaCl, 2% (w/v) glycerol, 0.01% (w/v) DDM, 0.01% (w/v) CHS and 20 mM imidazole] for 3 h at 4 C

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Mizuguchi Y, Sakamoto T, Hashimoto T, Tsukamoto S, Iwasa S, Saito Y, et al