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The (His) 6 -tag was removed by overnight thrombin cleavage, followed by polishing with size-exclusion chromatography (SEC) using a Superdex 75 HiLoad 16/600 column (Cytiva, MA, USA) equilibrated in 20 mM Tris-HCl pH 8.0, and 150 mM NaCl

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Gradual weight loss, under medical supervision, minimizes this risk

Reported side effects are rare and typically limited to mild irritation at the injection site or transient flu-like symptoms in the first few days