p53 is a tumour suppressor gene that has been explored for cancer gene therapy as a possible alternative to the common treatments. The use of plasmid DNA (pDNA) to carry the therapeutic gene has been considered, but it is requisite to preserve its supercoiled (sc) structure, for eliciting a more effective gene expression and therapeutic action. The purification of the sc pDNA using amino acids-based affinity chromatography has been successfully applied, exploring different amino acids and supports. From these studies, it stood out the selectivity of arginine for the recognition of sc pDNA. However, some limitation on the binding capacity was found in the arginine-agarose support, and in the case of monoliths, some fouling and clogging can limit sequential runs. By using macroporous support modified with arginine it was expected to take advantage of the selectivity of the ligand combined with the flow properties and binding capacity offered by the support. The arginine-modified macroporous support was characterized by SEM, EDX and FTIR also to verify the correct immobilization of arginine, and then used for pDNA purification. The support showed to be effective on the sc p53-pDNA isolation, and the robustness was also achieved by accomplishing the purification of plasmids with different sizes, only by slightly adjusting the experimental conditions. Regarding the dynamic binding capacity of the arginine-modified macroporous support, it was achieved an improvement of more than 50% in the pDNA binding capacity when compared with their homologous arginine-agarose commercial matrix, suggesting potential economic feasibility in case of scale-up.

中文翻译：

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使用精氨酸修饰的大孔支持物纯化超螺旋p53编码质粒。

p53是一种抑癌基因，已经被研究用于癌症基因治疗，可以作为常见治疗方法的替代方法。已经考虑使用质粒DNA（pDNA）携带治疗基因，但是必须保持其超螺旋（sc）结构，以引起更有效的基因表达和治疗作用。使用基于氨基酸的亲和层析纯化sc pDNA已成功应用，探索了不同的氨基酸和支持物。从这些研究中，可以看出精氨酸对sc pDNA的选择性。然而，在精氨酸-琼脂糖载体中发现了对结合能力的一些限制，并且在整料的情况下，一些结垢和堵塞会限制顺序运行。通过使用精氨酸修饰的大孔载体，期望利用配体的选择性以及载体提供的流动性和结合能力。用SEM，EDX和FTIR对精氨酸修饰的大孔载体进行了表征，以验证精氨酸的正确固定，然后用于pDNA纯化。该载体显示出对sc p53-pDNA分离有效，并且仅通过稍微调整实验条件即可完成具有不同大小的质粒的纯化，从而获得了鲁棒性。关于精氨酸修饰的大孔支持物的动态结合能力，与它们的同源精氨酸-琼脂糖商业基质相比，pDNA结合能力提高了50％以上，