Influenza virus is a huge financial and social burden for health care systems over the world. Currently, traditional approaches are not effective in the fight of the epidemy and new alternatives like DNA vaccines have been developed. However, the downstream process of DNA vaccines is a constant challenge in the biotechnology industry. Cryogels has several advantages over traditional supports and have been tested as stationary phase in chromatographic separations. In this work, a method based on poly(2-hydroxyethyl methacrylate) cryogel was used to purify the plasmid NTC7482-41H-VA2 HA, which express the Influenza hemagglutinin gene. For this purpose, the cryogel was synthesized by cryo-polymerization of 2-hydroxyethyl methacrylate and characterized by scanning electron microscopy. The purification of supercoiled isoform of the plasmid NTC7482-41H-VA2 HA from a clarified lysate sample was achieved in a two-step experiment using NaCl and the dynamic binding capacity of pHEMA cryogel was determined. The assessment of DNA vaccine allowed to conclude that the level of contaminants such as proteins, genomic DNA, RNA and endotoxins are in accordance with FDA agency.

流感病毒是全世界医疗保健系统的巨大财务和社会负担。当前，传统方法在对抗流行病方面无效，并且已经开发了诸如DNA疫苗的新替代品。但是，DNA疫苗的下游工艺在生物技术行业中一直是一个挑战。与常规载体相比，Cryogels具有多个优点，并且已在色谱分离中作为固定相进行了测试。在这项工作中，使用了一种基于聚（甲基丙烯酸2-羟乙酯）冷冻凝胶的方法来纯化表达流感血凝素基因的质粒NTC7482-41H-VA2 HA。为此目的，通过对甲基丙烯酸2-羟乙酯进行冷冻聚合来合成冷冻凝胶，并通过扫描电子显微镜对其进行表征。使用NaCl通过两步实验从澄清的裂解物中纯化质粒NTC7482-41H-VA2 HA的超螺旋同工型，并测定pHEMA冷冻凝胶的动态结合能力。对DNA疫苗的评估可以得出结论，即蛋白质，基因组DNA，RNA和内毒素等污染物的水平符合FDA机构的规定。