College of Chemistry and Chemical Engineering Central South University, Changsha, Hunan 410083, PR China; Hunan Provincial Key Laboratory of Efficient and Clean Utilization of Manganese Resources, Central South University, Changsha, Hunan 410083, PR China.
College of Chemistry and Chemical Engineering Central South University, Changsha, Hunan 410083, PR China.
Talanta. 2018 Sep 1;187:142-147. doi: 10.1016/j.talanta.2018.05.009. Epub 2018 May 4.
The accurate quantification of p53 protein expression level is of great importance for cancer diagnosis. Here, a highly sensitive fluorescent sensor based on DNA functionalized magnetic nanoparticles was developed for the detection of p53 protein expression. Instead of a monoclonal antibody, a consensus DNA was employed to capture p53 protein. Meanwhile the fluorescent dye tethered DNA was used as the signal output instead of enzyme tagged nanoparticle or antibody. Consequently, our developed method is cost-effective for both the p53 capture and detection by compared with the conventional immunoassay. The biosensor developed by the above strategy was used to quantitatively detect p53, which yields a detection limit of 8 p.M. with the linear range of 50 p.M. to 2 nM. The sensitive for specific p53 detection was achieved due to the facile magnetic separation from the complex condition, and the reduced non-specific absorption effect by dextran. Moreover, the method is able to measure p53 from real cell lysate without extensive sample pretreatment/separation. The developed p53 biosensor has high sensitivity, good selectivity and reliable accuracy. It demonstrates great potential in clinical cancer diagnosis and early detection of cancer.
准确量化 p53 蛋白表达水平对癌症诊断具有重要意义。在这里，开发了一种基于 DNA 功能化磁性纳米粒子的高灵敏度荧光传感器，用于检测 p53 蛋白的表达。该传感器采用共识 DNA 代替单克隆抗体来捕获 p53 蛋白。同时，荧光染料连接的 DNA 被用作信号输出，而不是酶标记的纳米粒子或抗体。因此，与传统的免疫测定相比，我们开发的方法在 p53 捕获和检测方面都具有成本效益。通过上述策略开发的生物传感器用于定量检测 p53，其检测限为 8 p.M，线性范围为 50 p.M 至 2 nM。由于从复杂条件中易于进行磁性分离，并通过葡聚糖减少了非特异性吸附效应，因此实现了针对特异性 p53 检测的高灵敏度。此外，该方法无需广泛的样品预处理/分离即可测量真实细胞裂解物中的 p53。所开发的 p53 生物传感器具有高灵敏度、良好的选择性和可靠的准确性。它在癌症的临床诊断和早期检测方面具有巨大的潜力。
