Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu 273165, People's Republic of China.
Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu 273165, People's Republic of China; Key Laboratories of Chemical Safety and Health, China National Center for Food Safety Risk Assessment, Beijing 100050, People's Republic of China.
Biosens Bioelectron. 2016 Nov 15;85:358-362. doi: 10.1016/j.bios.2016.05.021. Epub 2016 May 6.
Early detection and diagnosis have great practical significances for the effective prevention and treatment of cancer. In this study, we developed a novel, facile and ultra-sensitive fluorescence assay for the determination of tumor invasive biomarker β-glucuronidase (GLU) based on the inner-filter effect (IFE). The nitrogen-doped carbon quantum dots (N-CQDs) with green photoluminescence were employed as the fluorophore in IFE, and 4-nitrophenyl-β-D-glucuronide (PNPG) was used to act as GLU substrate, and GLU catalytic product (p-nitrophenol (PNP)) was capable of acting as the robust absorber in IFE to turn off the fluorescence of N-CQDs due to the complementary overlap between the absorption of PNP and the excitation of N-CQDs. Thus, signal of GLU activity could be recorded by the fluorescence intensity of N-CQDs. Unlike other fluorescence sensing mechanism such as fluorescence resonance energy transfer (FRET) or photoinduced electron transfer (PET), IFE has no requirement for electron or energy transfer process or any chemical modification of fluorophore, which makes our assay more flexible and simple. The proposed method exhibited a good linear relationship from 1UL(-1) to 60UL(-1) (R(2)=0.9967) with a low detection limit of 0.3UL(-1). This method was also successfully applied to the analysis of serum samples and the inhibitor screening from natural product. The developed sensor platform was proven to be reliable, facile, sensitive, and selective, making it promising as a candidate for GLU activity detection in clinic tumor diagnose and anti-tumor drug screening.
早期发现和诊断对于癌症的有效预防和治疗具有重要的实际意义。在这项研究中,我们基于内滤效应(IFE)开发了一种新颖、简便、超灵敏的荧光测定法,用于测定肿瘤侵袭性生物标志物β-葡糖苷酸酶(GLU)。具有绿色光致发光的氮掺杂碳量子点(N-CQDs)被用作IFE中的荧光团,4-硝基苯-β-D-葡糖苷酸(PNPG)被用作 GLU 底物,GLU 催化产物(对硝基苯酚(PNP))能够作为IFE 中的强吸收剂,由于 PNP 的吸收与 N-CQDs 的激发之间的互补重叠,从而关闭 N-CQDs 的荧光。因此,可以通过 N-CQDs 的荧光强度记录 GLU 活性的信号。与其他荧光传感机制(如荧光共振能量转移(FRET)或光诱导电子转移(PET))不同,IFE 不需要电子或能量转移过程或荧光团的任何化学修饰,这使得我们的测定法更加灵活和简单。该方法在 1UL(-1)至 60UL(-1)范围内呈现良好的线性关系(R(2)=0.9967),检测限低至 0.3UL(-1)。该方法还成功应用于血清样品的分析和天然产物的抑制剂筛选。所开发的传感器平台被证明可靠、简便、灵敏且具有选择性,有望成为临床肿瘤诊断和抗肿瘤药物筛选中 GLU 活性检测的候选方法。