Cheng Na, Liu Yujie, Mukama Omar, Han Xiaobo, Huang Hualin, Li Shuai, Zhou Peng, Lu Xuewen, Li Zhiyuan
Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University Changsha China
Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences Guangzhou 510530 China.
RSC Adv. 2020 May 18;10(32):18601-18607. doi: 10.1039/d0ra02662j. eCollection 2020 May 14.
Platelet-derived growth factor BB (PDGF-BB) is a potential biomarker of tumor angiogenesis. For the first time, we developed a highly sensitive aptasensor for PDGF-BB with an enhanced test line signal by using two different gold nanoparticles (AuNPs). Herein, we describe a highly sensitive biosensor for PDGF-BB detection that combines biotinylated aptamer on a sample pad and poly thymine-Cy3-AuNP-monoclonal antibody complexes against PDGF-BB immobilized on conjugate pad A. Streptavidin (SA) and rabbit anti-mouse polyclonal antibody were also immobilized in the nitrocellulose membrane at the test and control zones, respectively. When the target PDGF-BB protein was added, it first bound the aptamer, and later the monoclonal antibody to form a biotinylated complex that was captured by SA, resulting in a visual red line on the test zone. In addition, to enhance the sensitivity, another monoclonal antibody against Cy3 was conjugated on AuNP B and immobilized on conjugate pad B to form a AuNPs (A&B)-antibody-(PDGF-BB-Cy3)-aptamer-biotin-SA complex on the test line when a loading buffer was subsequently added. This approach showed a linear response to PDGF-BB from 3 ng mL to 300 ng mL with a limit of detection as low as 1 ng mL obtained in 10 minutes. Our biosensor displayed results through red lines readable by the naked eye. Interestingly, our approach has been successfully applied for real sample verification, proving its applicability for cancer monitoring and diagnosis.
血小板衍生生长因子BB(PDGF-BB)是肿瘤血管生成的一种潜在生物标志物。我们首次开发了一种用于PDGF-BB的高灵敏度适配体传感器,通过使用两种不同的金纳米颗粒(AuNP)增强检测线信号。在此,我们描述了一种用于检测PDGF-BB的高灵敏度生物传感器,该传感器将生物素化适配体置于样品垫上,并将针对PDGF-BB的聚胸腺嘧啶-Cy3-AuNP-单克隆抗体复合物固定在共轭垫A上。链霉亲和素(SA)和兔抗小鼠多克隆抗体也分别固定在硝酸纤维素膜的检测区和对照区。当加入目标PDGF-BB蛋白时,它首先与适配体结合,随后与单克隆抗体形成生物素化复合物,该复合物被SA捕获,在检测区产生可见的红线。此外,为了提高灵敏度,将另一种抗Cy3单克隆抗体与AuNP B偶联并固定在共轭垫B上,当随后加入加样缓冲液时,在检测线上形成AuNPs(A&B)-抗体-(PDGF-BB-Cy3)-适配体-生物素-SA复合物。该方法对PDGF-BB的线性响应范围为3 ng/mL至300 ng/mL,10分钟内检测限低至1 ng/mL。我们的生物传感器通过肉眼可读的红线显示结果。有趣的是,我们的方法已成功应用于实际样品验证,证明了其在癌症监测和诊断中的适用性。