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3.5 Removal of GST affinity tag after protease cleavage in solution If the protein was digested in solution, the target protein can be purified further by re-chromatography on the glutathione Sepharose column to remove any un-cleaved fusion protein and the GST moiety

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the ligand is a ligand that is endocytosed and/or transcytosed into and/or through the target cell as may be assayed by a cell culture model of the BBB (using primary isolated bovine brain capillary endothelial cells (BCEC)) as described by Gaillard et al