Abstract：
Plasmids are essential source material for production of biological drugs, vaccines and vectors for gene therapy. They are commonly formulated as frozen solutions. Considering the cost associated with maintenance of cold chain conditions during storage and transport, there is a significant need for alternative methods for stabilization of plasmids at ambient temperature. The objective of these studies was to identify a film-based formulation that preserved transfection efficiency of plasmids at 25 °C. A model plasmid, pAAVlacZ, was used for these studies. Transfection efficiency and agarose gel electrophoresis were utilized to assess bioactivity and changes in physical conformation of plasmid during storage. An amino acid, capable of sustaining a positive charge while supporting an alkaline environment within the film matrix, preserved transfection efficiency for 9 months at 25 °C. Addition of sugar and a plasticizer to the formulation preserved the plasmid in an amorphous state and improved handling properties of the film. The manner in which excipients were incorporated into bulk formulations and environmental humidity in which films were stored significantly impacted transfection efficiency of plasmid in the rehydrated solution. Taken together, these results suggest that plasmids can be stored for extended periods of time without refrigeration within a film matrix.
摘要：
质粒是生物药物生产的重要原料,基因治疗的疫苗和载体。它们通常配制成冷冻溶液。考虑到储存和运输过程中与维护冷链条件相关的成本，对于在环境温度下稳定质粒的替代方法存在显著的需要。这些研究的目的是鉴定在25°C下保持质粒转染效率的基于膜的制剂。一个模型质粒，pAAVlacZ,用于这些研究。转染效率和琼脂糖凝胶电泳用于评估生物活性和储存过程中质粒物理构象的变化。一种氨基酸，能够维持正电荷，同时支持薄膜基质内的碱性环境，在25°C下保持9个月的转染效率向制剂中添加糖和增塑剂使质粒保持在无定形状态并改善膜的处理性质。将赋形剂掺入散装制剂中的方式和储存膜的环境湿度显著影响质粒在再水合溶液中的转染效率。一起来看,这些结果表明，质粒可以在薄膜基质中长时间储存而无需冷藏。
