Human acidic fibroblast growth factor (haFGF) is a multifunctional protein involved in regulating a wide range of cellular processes. As a potent therapeutic agent, it is highly desirable to produce recombinant haFGF (r-haFGF) at low cost. However, the complex structure and formation of aggregation confines its high-level soluble expression and functional form. Herein, to produce r-haFGF efficiently in E. coli, we devised a novel soluble expression and cost-effective purification approach based on fusion with Scl2-M (a novel modified collagen-like protein) for the first time. By using this strategy, more than 95% of the Scl2-M-haFGF fusion protein was highly expressed in soluble form and the expression level of targeted fusion protein in shake flasks and 5-L fermenter was 0.42 g/L and 2.28 g/L, respectively. Subsequently, the recombinant Scl2-M-haFGF was readily purified through a facile process of acid precipitation and subjected to enterokinase (EK) cleavage. After Scl2-M cleavage, tag-free r-haFGF was further purified using ion-exchange chromatography. The recovery rate of the whole purification process attained 34.2%. Furthermore, the resulting high-purity (96.0%) r-haFGF was prepared by freeze-drying as a final product, and its bioactivity was confirmed to potentiate the proliferation of L929 and BALB-3T3 fibroblasts. Overall, our developed method has the potential for the massive production of the r-haFGF in the future.

人酸性成纤维细胞生长因子（haFGF）是一种多功能蛋白，参与调节广泛的细胞过程。作为有效的治疗剂，非常希望以低成本生产重组haFGF（r-haFGF）。然而，聚集体的复杂结构和形成限制了其高水平的可溶性表达和功能形式。本文中，为了在大肠杆菌中有效产生r-haFGF，我们首次设计了一种基于与Scl2-M（一种新型修饰的胶原蛋白样蛋白）融合的新型可溶性表达和具有成本效益的纯化方法。通过使用该策略，超过95％的Scl2-M-haFGF融合蛋白以可溶性形式高表达，并且在摇瓶和5-L发酵罐中目标融合蛋白的表达水平为0.42 g / L和2.28 g / L ， 分别。随后，通过酸沉淀的简便方法容易地纯化重组Scl2-M-haFGF，并进行肠激酶（EK）裂解。Scl2-M裂解后，使用离子交换色谱法进一步纯化无标签的r-haFGF。整个纯化过程的回收率达到34.2％。此外，通过冷冻干燥作为最终产品来制备所得的高纯度（96.0％）r-haFGF，并证实其生物活性可增强L929和BALB-3T3成纤维细胞的增殖。总的来说，我们开发的方法有可能在未来大量生产r-haFGF。