Table 2.

Approximate time from overexpression to obtaining pure recombinant target protein

Subprocess	Approximate time required
Bacterial transformation	2 h
Growth of transformants on plate	12–16 h
Colony pickup and growth of starter culture	12–16 h
Large-scale expression, induction, and harvesting	8 h
Sonication, centrifugation, and protein purification	8 h
Buffer exchange/desalting of the desired protein	2 h
Enzymatic cleavage	24 h
Heat treatment	20 min
Separation of GST tag from the target protein using high-speed centrifugation	20 min