Table 3. Common proteases used for fusion tag removal.
Protease | Advantages | Disadvantages |
---|---|---|
TEV | Stringent cleavage-sequence specificity and few residual amino acids after cleave. Easily produced in house. Good activity in a range of buffers and at 4°C. |
Activity limited by some commonly used detergents. Reducing agents required for activity. Comparatively low activity. |
HRV 3C | Stringent cleavage-sequence specificity and few residual amino acids after cleave. Easily produced in house. Good activity in a range of buffers and at 4°C. |
Activity limited by some commonly used detergents. Comparatively low activity. |
Thrombin | Not affected by the majority of detergents. | Non-specific cleavage, Inhibited by reducing agents and common protease inhibitors used during purification. |
SUMO protease | No recombinant linker region needs to be constructed, native N-terminus of the target protein is maintained. | Little activity in many commonly used detergents. |
Factor Xa | Not affected by the majority of detergents. | Non-specific cleavage, Inhibited by reducing and chelating agents, phosphate ions. |