Department of Chemistry in Pharmaceutical Sciences, Faculty of Pharmacy , Complutense University of Madrid , Plaza Ramon y Cajal, No. 2 , 28040 , Madrid , Spain.
Department of Biotechnology , University of Turku , Kiinamyllynkatu 10 , FI-20520 Turku , Finland.
Anal Chem. 2018 Nov 20;90(22):13385-13392. doi: 10.1021/acs.analchem.8b03106. Epub 2018 Oct 31.
In this work, we explore a photochemical ligation reaction to covalently modify oligonucleotide-conjugated upconverting nanoparticles (UCNPs) in the presence of a specific target DNA sequence. The target sequence acts as a hybridization template, bringing together a biotinylated photoactivatable oligonucleotide probe and the oligonucleotide probe that is attached to UCNPs. The illumination of the UCNPs by NIR light to generate UV emission internally or illuminating the photoactivatable probe directly by an external UV light promotes the photochemical ligation reaction, yielding covalently biotin functionalized UCNPs that can be selectively captured in streptavidin-coated microwells. Following this strategy, we developed a DNA sensor with a limit of detection of 1 × 10 mol per well (20 fM). In addition, we demonstrate the possibility to create UCNP patterns on the surface of solid supports upon NIR illumination that are selectively formed under the presence of the target oligonucleotide.
在这项工作中，我们探索了一种光化学连接反应，以在特定目标 DNA 序列存在的情况下，将寡核苷酸缀合的上转换纳米粒子 (UCNP) 进行共价修饰。目标序列充当杂交模板，将生物素化的光活化寡核苷酸探针和连接到 UCNP 上的寡核苷酸探针聚集在一起。通过近红外光照射 UCNP 以在内部产生紫外发射，或直接通过外部紫外光照射光活化探针，促进光化学连接反应，产生共价生物素化的 UCNP，其可以在链霉亲和素包被的微孔中被选择性捕获。根据这一策略，我们开发了一种 DNA 传感器，其检测限为每孔 1×10-12 mol（20 fM）。此外，我们证明了在近红外光照射下，可以在固体载体表面上选择性地形成 UCNP 图案，并且只有在存在目标寡核苷酸的情况下才会形成这些图案。
