Zhang Jing-Jing, Cao Jun-Tao, Shi Gui-Fang, Liu Yan-Ming, Chen Yong-Hong, Ren Shu-Wei
College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000, China.
Talanta. 2015 Aug 15;141:158-63. doi: 10.1016/j.talanta.2015.04.001. Epub 2015 Apr 9.
This work describes a novel electrochemiluminescence aptasensor for highly sensitive detection of platelet-derived growth factor BB (PDGF-BB) using aptamer functionalized CdS quantum dots-polyamidoamine as probe (CdS QDs-PAMAM-Apt). CdS QDs-PAMAM nanocomposites were synthesized by one-pot synthesis in methanol. The prepared nanocomposites were linked with the NH2-aptamer 2 (Apt2) of PDGF-BB to form the CdS QDs-PAMAM-Apt2 probe by glutaraldehyde as coupling reagent. For constructing the aptasensor, MWCNTs-chitosan composites and NH2-aptamer 1 (Apt1) with the same base sequence as Apt2 were immobilized on the electrode by the self-assembled method to recognize the target protein PDGF-BB. In the presence of PDGF-BB, the structure of sandwiched format was formed between the Apt1 and the CdS QDs-PAMAM-Apt2 probe, thereby resulting in a proportional increase of ECL emission. Thanks to the efficient and stable ECL emission of CdS QDs-PAMAM dendrimer and the advantage of MWCNTs for accelerating the electron transfer, the highly sensitive detection of PDGF-BB with a detection limit of 0.13pM was achieved. The linear range is from 0.5pM to 1nM. The present protocol was applied to the analysis of PDGF-BB in human serum samples. The recoveries of PDGF-BB in human serum samples are 87.2-113% and RSD values are less than 3.6%.
本研究报道了一种新型的电化学发光适体传感器,该传感器以适体功能化的硫化镉量子点-聚酰胺胺为探针(CdS QDs-PAMAM-Apt),用于高灵敏度检测血小板衍生生长因子BB(PDGF-BB)。CdS QDs-PAMAM纳米复合材料通过在甲醇中一锅法合成。制备的纳米复合材料通过戊二醛作为偶联剂与PDGF-BB的NH2-适体2(Apt2)连接,形成CdS QDs-PAMAM-Apt2探针。为构建适体传感器,将具有与Apt2相同碱基序列的多壁碳纳米管-壳聚糖复合材料和NH2-适体1(Apt1)通过自组装方法固定在电极上,以识别目标蛋白PDGF-BB。在存在PDGF-BB的情况下,在Apt1和CdS QDs-PAMAM-Apt2探针之间形成夹心结构,从而导致电化学发光发射成比例增加。由于CdS QDs-PAMAM树枝状大分子具有高效稳定的电化学发光发射以及多壁碳纳米管在加速电子转移方面的优势,实现了对PDGF-BB的高灵敏度检测,检测限为0.13pM。线性范围为0.5pM至1nM。本方法应用于人血清样品中PDGF-BB的分析。人血清样品中PDGF-BB的回收率为87.2-113%,相对标准偏差值小于3.6%。