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Table 3 Conditions for solubilization of inclusion bodies reported in human insulin/analogue downstream processing, ranked in order of ascending duration of solubilization

From: Downstream processing of recombinant human insulin and its analogues production from E. coli inclusion bodies

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)
  1. NA information not available, RTP room temperature