Refolding Record:
| Protein | |
|---|---|
| Protein Name | Interferon gamma |
| Abbreviated Name | IFN gamma |
| SCOP Family | Interferons/interleukin-10 (IL-10) |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01579 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 140 |
| Molecular Weight | 16177.5 |
| Pi | 9.52198 |
| Molecular Weight | 16177.5 |
| Disulphides | 0 |
| Full Sequence |
QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWK
EESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTN
YSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRG
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Gong B, Wang L, Wang C, Geng X (2004) Journal of Chromatography A, 1022, 33-39 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | DH5? |
| Expression Temp | 37.0 |
| Expression Time | 12h |
| Expression Vector | pBV 220 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | Lysozyme |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: hydrophobic interaction chromatography |
| Wash Buffer | 20 mM PBS |
| Solubilization Buffer | 7.0 M GuHCl |
| Refolding Buffer | 50 mM PBS |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no tag |
| Refolding pH | 7.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | 45 min |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The inclusion bodies of rhIFN- were disrupted with buffer consisting of 20 mmol/l phosphate + 1 mmol/l EDTA + 0.2 mg/ml lysozyme (pH = 7.4), then the inclusion bodies were washed three times. Finally, the inclusion bodies were dissolved in 7.0 mol/l guanidine hydrochloride (Gu.HCl) solution. After incubation at 4 ?C for 24 h with full agitation, the supernatant of rhIFN- was obtained by centrifuging it at 20000 rpm. A 5.0 × 0.8 cm i.d. HIC column was used to purification and simultaneous renaturation of rhIFN- in the extract solution. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | 95% |
| Notes | |