当前位置：
X-MOL 学术
›
ACS Comb. Sci.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Protein-Induced Fluorescence Enhancement Based Detection of Plasmodium falciparum Glutamate Dehydrogenase Using Carbon Dot Coupled Specific Aptamer
ACS Combinatorial Science Pub Date : 2018-05-03 00:00:00 , DOI: 10.1021/acscombsci.8b00021 Naveen Kumar Singh 1 , Babina Chakma 1 , Priyamvada Jain 1 , Pranab Goswami 1
ACS Combinatorial Science Pub Date : 2018-05-03 00:00:00 , DOI: 10.1021/acscombsci.8b00021 Naveen Kumar Singh 1 , Babina Chakma 1 , Priyamvada Jain 1 , Pranab Goswami 1
Affiliation
|
A novel 90-mer long ssDNA aptamer (NG3) covering a 40-mer random region targeting Plasmodium falciparum glutamate dehydrogenase (PfGDH) developed through systematic evolution of ligands by exponential enrichment (SELEX) technique. The binding affinity of the aptamer to PfGDH discerned by circular dichroism (CD) was 0.5 ± 0.04 μM. The specificity of the aptamer toward the target was confirmed by gel electrophoresis and CD studies. The presence of two quadruplex forming regions, two big and four small stem loop structures with a δG of −7.99 kcal mol–1 for NG3 were deduced by computational studies. The spherical carbon dots (Cdots) of size 2–4 nm, synthesized by pyrolysis method using l-glutamate as a substrate were covalently linked to the amine modified aptamer. The Cdot with a band gap of 2.8 eV and a quantum yield of 34% produced fluorescence at ∼ λ410 nm when excited at λ320nm. The quantum yield of Cdot-aptamer assembly was increased up to 40% in the presence of the PfGDH in solution. A linear relationship with a dynamic range of 0.5 nM to 25 nM (R2 = 0.98) and a limit of detection (LOD) of 0.48 nM was observed between the fluorescence intensity of the Cdots-aptamer conjugate and the concentration of PfGDH. The method could detect PfGDH with an LOD of 2.85 nM in diluted serum sample. This novel simple, sensitive and specific protein induced fluorescence enhancement based detection of PfGDH has a great potential to develop as a method for malaria detection.
中文翻译：
碳点偶联特异性适体基于蛋白质诱导的荧光增强的恶性疟原虫谷氨酸脱氢酶检测
新型的90-mer长ssDNA适体（NG3）覆盖了40-mer靶向恶性疟原虫谷氨酸脱氢酶（Pf GDH）的随机区域，是通过配体通过指数富集（SELEX）技术进行系统进化而开发的。通过圆二色性（CD）识别的适体对Pf GDH的结合亲和力为0.5±0.04μM。通过凝胶电泳和CD研究证实了适体对靶的特异性。通过计算研究推导了存在两个四联体形成区域，两个大和四个小的茎环结构，其NG3的δG为-7.99 kcal mol –1。使用l通过热解法合成的大小为2-4 nm的球形碳点（Cdots）-谷氨酸作为底物与胺修饰的适体共价连接。带隙为2.8 eV且量子产率为34％的Cdot在λ320nm激发时在λ410 nm处产生荧光。在溶液中存在Pf GDH的情况下，Cdot-适体组装体的量子产率提高至40％。在Cdots-适体缀合物的荧光强度和Pf GDH的浓度之间观察到动态范围为0.5 nM至25 nM（R 2 = 0.98）和检测极限（LOD）为0.48 nM的线性关系。该方法可以检测Pf稀释的血清样品中LOD为2.85 nM的GDH。这种新颖，简单，灵敏和特异的蛋白质诱导的基于荧光增强的Pf GDH的检测方法具有巨大的潜力，可以作为一种检测疟疾的方法。
更新日期：2018-05-03
中文翻译：
碳点偶联特异性适体基于蛋白质诱导的荧光增强的恶性疟原虫谷氨酸脱氢酶检测
新型的90-mer长ssDNA适体（NG3）覆盖了40-mer靶向恶性疟原虫谷氨酸脱氢酶（Pf GDH）的随机区域，是通过配体通过指数富集（SELEX）技术进行系统进化而开发的。通过圆二色性（CD）识别的适体对Pf GDH的结合亲和力为0.5±0.04μM。通过凝胶电泳和CD研究证实了适体对靶的特异性。通过计算研究推导了存在两个四联体形成区域，两个大和四个小的茎环结构，其NG3的δG为-7.99 kcal mol –1。使用l通过热解法合成的大小为2-4 nm的球形碳点（Cdots）-谷氨酸作为底物与胺修饰的适体共价连接。带隙为2.8 eV且量子产率为34％的Cdot在λ320nm激发时在λ410 nm处产生荧光。在溶液中存在Pf GDH的情况下，Cdot-适体组装体的量子产率提高至40％。在Cdots-适体缀合物的荧光强度和Pf GDH的浓度之间观察到动态范围为0.5 nM至25 nM（R 2 = 0.98）和检测极限（LOD）为0.48 nM的线性关系。该方法可以检测Pf稀释的血清样品中LOD为2.85 nM的GDH。这种新颖，简单，灵敏和特异的蛋白质诱导的基于荧光增强的Pf GDH的检测方法具有巨大的潜力，可以作为一种检测疟疾的方法。
京公网安备 11010802027423号