Fusion protein | Media | Comments | References |
---|---|---|---|
Denatured poly-histidine/ PI fusion protein | Ni2+–activated chelating Sepharose | Washed with buffer A: 20 mM NaPO4, pH 7.4, 500 mM NaCl, 20 mM imidazole, 8 M urea Elution with linear gradient (10 CVs), ending with buffer A containing 500 mM imidazole | Mackin (1999) |
Histidine-tagged denatured PI | Ni–NTA His•Bind Superflow | Capture Histidine-tagged peptide Binding and elution buffers contain 8 M urea Elution with 0.15 M imidazole, pH 7.5 Yield: 77.8%; Purity: > 79% → 97.6% | Yuan et al. (2015) |
Histidine-tagged sulfonated PI | Ni-chelating Sepharose FF | Stepwise gradient elution with 8 M urea and 0.08 M imidazole | Astolfi et al. (2004) |
Histidine-tagged sulfonated PI | NTA column | Eluted according to a standard protocol (Qiaexpression, Qiagen) | Redwan et al. (2007) |
Hexahistidine-tagged sulfonated PI | Ni-IDA-Sepharose | Binding and elution buffers contain 6 M urea Elution with 0.1 M imidazole Yield: 90% Purity: 85–95% (order of chromatography affects purity) High selectivity to poly-His sequence | Tikhonov et al. (2001) |
Histidine-tagged renatured PI | IMAC | Elution with a 15 CV gradient from 0 to 400Â mM imidazole Purity: 92% | Zimmerman and Stokell (2010) |
Z-PI (secreted) | IgG Sepharose 6 Fast Flow | Equilibration with Tris-Saline-Tween (TST) buffer (1 mM EDTA 25 mM Tris–HCl, pH 8, 0.2 M NaCl, 0.5 mL Tween 20) Elution with 0.5 mM acetic acid, pH 2.8 | Mergulhao et al. (2004) |
ZZ-PI (secreted) | IgG Sepharose 6 Fast Flow | Equilibration with Tris-Saline-Tween (TST) buffer (1 mM EDTA 25 mM Tris–HCl, pH 8, 0.2 M NaCl, 0.5 mL Tween 20) Elution with 0.5 mM acetic acid, pH 2.8 | Mergulhao et al. (2001) |
ZZ-PI | IgG Sepharose | Binding buffer contains 0.1 M glycine–NaOH, BME Elution with 0.3 M acetic acid, pH 3.1 Yield: 90%  ~ 70% of ZZ-R-proinsulin was recovered in monomeric form | Nilsson et al. (1996) |