Cao Zhi-Juan, Peng Qian-Wen, Qiu Xue, Liu Cai-Yun, Lu Jian-Zhong
School of Pharmacy, Fudan University, Shanghai 201203, China.
J Pharm Anal. 2011 Aug;1(3):159-165. doi: 10.1016/j.jpha.2011.06.002. Epub 2011 Jul 22.
A robust, selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.
本文提出了一种用于蛋白质检测的稳健、选择性高且灵敏度高的化学发光（CL）平台。这种新型CL方法利用滚环扩增（RCA）作为信号增强技术，并将96孔板用作固定和分离载体。通常，固定在96孔板表面的抗体与蛋白质靶标和适体-引物序列夹在一起。然后将该适体-引物序列用作RCA的引物。基于此设计，大量生物素化探针和链霉亲和素-辣根过氧化物酶（SA-HRP）被捕获在板上，CL信号被放大。总之，我们的结果证明了一种稳健的生物传感器，其检测限为10 fM，易于建立和使用，且无需光源。因此，这项新技术将利用其高灵敏度和选择性，拓宽基于DNA的生物传感器未来用于检测与临床疾病相关的其他蛋白质生物标志物的发展前景。
