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Comprehensive enzymatic tag-removal reviews highlight TEV’s advantages over other proteases—including broad N-terminal residue tolerance, mild-temperature activity (including at 4 °C), and robust cleavage efficiency, while also pointing out limitations such as occasional autolysis (that can be mitigated by S219V mutation in the TEV enzyme), reduced activity at higher temperatures, and the need for reducing agents in some constructs [139,161,162]

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