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To gain insight into the nature of the hydroxocobalamin-mAb binding, a portion of the DTT-containing mAb sample was alkylated with 0.25 M iodoacetic acid (I4386–10G, Sigma Aldrich) prior to buffer exchange to create an irreversibly reduced mAb sample with covalently modified thiol groups

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B12 is water-soluble, so the body generally excretes what it does not need