K L E F University, Centre for Bioprocess Technology, Department of Biotechnology, Guntur 522 502, Andhra Pradesh, India.
Andhra University, College of Engineering, Visakhapatnam 530 003, Andhra Pradesh, India.
J Chromatogr A. 2015 Jan 30;1379:74-82. doi: 10.1016/j.chroma.2014.12.057. Epub 2014 Dec 29.
Protein folding-strong cation exchange chromatography (PF-SCX) has been employed for efficient refolding with simultaneous purification of recombinant human granulocyte colony stimulating factor (rhG-CSF). To acquire a soluble form of renatured and purified rhG-CSF, various chromatographic conditions, including the mobile phase composition and pH was evaluated. Additionally, the effects of additives such as urea, amino acids, polyols, sugars, oxidizing agents and their amalgamations were also investigated. Under the optimal conditions, rhG-CSF was efficaciously solubilized, refolded and simultaneously purified by SCX in a single step. The experimental results using ribose (2.0M) and arginine (0.6M) combination were found to be satisfactory with mass yield, purity and specific activity of 71%, ≥99% and 2.6×10(8)IU/mg respectively. Through this investigation, we concluded that the SCX refolding method was more efficient than conventional methods which has immense potential for the large-scale production of purified rhG-CSF.
蛋白折叠-强阳离子交换色谱（PF-SCX）已被用于高效复性，并可同时纯化重组人粒细胞集落刺激因子（rhG-CSF）。为了获得可溶形式的复性和纯化的 rhG-CSF，评估了各种色谱条件，包括流动相组成和 pH 值。此外，还研究了添加剂如尿素、氨基酸、多元醇、糖、氧化剂及其混合物的影响。在最佳条件下，rhG-CSF 可通过 SCX 在一步中有效溶解、复性和同时纯化。使用核糖（2.0M）和精氨酸（0.6M）组合的实验结果令人满意，质量产率、纯度和比活性分别为 71%、≥99%和 2.6×10(8)IU/mg。通过这项研究，我们得出结论，SCX 复性方法比传统方法更有效，对于大规模生产纯化的 rhG-CSF 具有巨大的潜力。
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