Nanoworld Institute; Fondazione EL.B.A. Nicolini; Pradalunga, Italy; Laboratories of Biophysics and Nanobiotechnology; Department of Experimental Medicine; University of Genova; Genova, Italy; Department of Surgical Sciences and Integrated Diagnostic; University of Genova; Genova, Italy.
Bioengineered. 2013 Sep-Oct;4(5):332-7. doi: 10.4161/bioe.25278. Epub 2013 Jun 12.
This paper investigates the application of anodic porous alumina as an advancement on chip laboratory for gene expressions. The surface was prepared by a suitable electrolytic process to obtain a regular distribution of deep micrometric holes and printed bypen robot tips under standard conditions. The gene expression within the Nucleic Acid Programmable Protein Array (NAPPA) is realized in a confined environment of 16 spots, containing circular DNA plasmids expressed using rabbit reticulocyte lysate. Authors demonstrated the usefulness of APA in withholding the protein expression by detecting with a CCD microscope the photoluminescence signal emitted from the complex secondary antibody anchored to Cy3 and confined in the pores. Friction experiments proved the mechanical resistance under external stresses by the robot tip pens printing. So far, no attempts have been made to directly compare APA with any other surface/substrate; the rationale for pursuing APA as a potential surface coating is that it provides advantages over the simple functionalization of a glass slide, overcoming concerns about printing and its ability to generate viable arrays.
本文研究了阳极多孔氧化铝在基因表达片上实验室中的应用。通过适当的电解过程制备表面,以获得深亚微米孔的规则分布,并在标准条件下使用笔式机器人尖端进行打印。在包含用兔网织红细胞裂解物表达的圆形 DNA 质粒的 16 个点的受限环境中实现了核酸可编程蛋白阵列 (NAPPA) 中的基因表达。作者通过检测固定在孔中的 Cy3 标记的二级抗体发出的光致发光信号,用 CCD 显微镜证明了 APA 在抑制蛋白表达方面的有用性。摩擦实验证明了机器人笔尖打印在外力作用下的机械阻力。到目前为止,尚未尝试直接将 APA 与任何其他表面/基底进行比较;选择 APA 作为潜在表面涂层的依据是,它提供了优于玻璃载玻片简单功能化的优势,克服了对打印及其生成可行阵列的能力的担忧。