The granulocyte-macrophage colony-stimulating factor (GM-CSF) can be used to induce a powerful immune response. Based on the specific binding of biotin and streptavidin, SA-hGM-CSF was anchored on the surface of biotinylated tumor cells, which could enhance the anti-tumor effect of tumor cell vaccines in our previous reports, suggesting it would have potential clinical value. Preparation of the biologically active proteins in large-scale production is the basis of clinical application, however, only a small amount of biologically active protein was obtained according to previous studies. In this study, we researched the effects of various factors on the purification and simultaneous renaturation of SA-hGM-CSF fusion protein by single factor experiment and orthogonal experiment. Here, we developed a viable pilot-scale trial in the fermentation, purification, refolding and freeze-drying of SA-hGM-CSF proteins in order to efficiently obtain more biologically active proteins with high purity, which will lay the foundation for industrial production.

粒细胞巨噬细胞集落刺激因子（GM-CSF）可用于诱导强大的免疫反应。基于生物素和链霉亲和素的特异性结合，SA-hGM-CSF被锚定在生物素化的肿瘤细胞表面，这可以增强我们先前报告中肿瘤细胞疫苗的抗肿瘤作用，表明其具有潜在的临床价值。大规模生产生物活性蛋白的制备是临床应用的基础，但是，根据先前的研究，仅获得了少量的生物活性蛋白。在这项研究中，我们通过单因素实验和正交实验研究了各种因素对SA-hGM-CSF融合蛋白的纯化和同时复性的影响。在这里，我们开发了可行的发酵试验规模，