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| Recombinant expressed human stem cell factor (rhSCF) as cytoplasmic inclusion bodies (IB) are reported. In present work, to increase mass recovery of rhSCF production in large scale, the factors affecting about efficiency of rhSCF IB were recovered and solubilised in urea solution, and refolding with simultaneous purification process using protein folding liquid chromatography (PFLC) were investigated, including normal chromatographic column, the unit for the simultaneous renaturation and purification of proteins (USRPP). Finally, by combining the optimized buffer and USRPP, we were able to obtain 22 mg rhSCF with >95% purity.The mass recovery is 24 % for dilution, 38 % for the normal chromatographic column, 49 % for the USRPP. An average specific bioactivity is 4.27 ×105 IU/mg, 6.9×105 IU/mg, and 1.28×106 IU/mg, respectively. These protocol dates and new refolding with purification method -USRPP provide a cost effective and an efficient way to produce quantities of high purity rhSCF in large-scale. |
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| Keywords:recombinant human stem cell factor, inclusion body, protein folding liquid chromatography |
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