Table 3 Conditions for solubilization of inclusion bodies reported in human insulin/analogue downstream processing, ranked in order of ascending duration of solubilization
Solubilization duration | Solubilization agent | Reducing agent | Chelating agent (EDTA) | Other components | Temp. (°C) | pH | Suspension concentration during solubilization (mg/mL) | References |
|---|---|---|---|---|---|---|---|---|
30 min | 8 M urea | – | – | – | NA | 8.9 | NA | Castellanos-Serra et al. (1996) |
30 min | – | – | 0.2 mM | 12 mM NaHCO3 | RTP | 12.0 | NA | Mikiewicz et al. (2017) |
45 min | – | – | 0.2 mM | 12 mM NaHCO3 | RTP | 11.9, then adjusted to 10.8 | NA | Zieliński et al. (2019) |
6 h | 8 M urea | – | 5 mM (optional) | 1% toluene (optional) | RTP | 8.5 | NA | Astolfi et al. (2004) |
8 h | 5 M urea | 40 g/L BME | – | – | NA | NA | NA | Petrides (1995) |
Overnight | 8 M urea | 3 mM BME | 1 mM | – | 4 | 8.0 | 100 | Bai et al. (2003) |
> 12 h | 8 M urea | 20 mM DTT | 1 mM | – | 4 | 8.0 | 100 | Yuan et al. (2015) |
NA | 6 M GdnHCl | 100 mM BME | – | – | NA | 8.5 | 167 | Chen et al. (2016) |
NA | 4 M urea | – | – | 10 mM glycine | NA | 10.6 | NA | Son et al. (2009) |
NA | 8 M urea | – | – | – | NA | 10.6 | NA | Hwang et al. (2016) |
NA | 8 M urea | – | 1 mM | – | NA | 11, then adjusted to 9.5 | 10–15 | Kim et al. (2015) |
NA | 8 M urea | 4 mM BME | – | – | NA | 10.4 | 20–30 | Leng et al. (2013) |
NA | 8 M urea | – | 1 mM | – | NA | 11, then adjusted to 9.5 | 10–15 | Min et al. (2011) |
NA | 8 M urea and 6 M GdnHCl | – | – | – | NA | 9.0 | 10 | Redwan et al. (2007) |
NA | 8 M GdnHCl | – | – | – | NA | 10.8 | NA | Thurow et al. (2010) |