Purification and Refolding of Overexpressed Human Basic Fibroblast Growth Factor in Escherichia coli

Figure 3

Purification of insoluble hbFGF with cation exchange expanded bed adsorption. Column: CM Hyper Z (60 mL of gel). Sample: unclarified cell lysate in 8 M urea, PBS pH 7. Equilibration buffer: 8 M urea, PBS pH 7. Refolding buffer: PBS pH 7. Elution buffer: 0.2–2 M NaCl, 20 mM sodium bicarbonate pH 9. Detection at 280 nm; flow rate: 40 mL/min. Flow through: no retained fraction; peak 1: 0.4 M NaCl eluted fraction; peak 2: 0.8 M NaCl eluted fraction.