Kim Chloe, Searson Peter C
Department of Materials Science and Engineering, Johns Hopkins University , 3400 North Charles Street, Baltimore, Maryland 21218, United States.
Institute for Nanobiotechnology at Johns Hopkins University , 3400 North Charles Street, Baltimore, Maryland 21218, United States.
Bioconjug Chem. 2017 Sep 20;28(9):2230-2234. doi: 10.1021/acs.bioconjchem.7b00328. Epub 2017 Aug 15.
To overcome the limitations associated with antibody-based sensors, we describe a proof-of-concept of an aptamer-based sandwich assay for detection of lactate dehydrogenase, an antigen associated with malaria. We show a detection limit of Plasmodium falciparum lactate dehydrogenase and Plasmodium vivax lactate dehydrogenase of 0.5 fmole in buffer, comparable to an antibody-based assay, using a magnetic particle-aptamer construct for capture and a quantum dot-aptamer construct for detection. We then demonstrate a detection limit of 10 amole (50-fold amplification) using oligonucleotide-functionalized gold nanoparticles to allow the conjugation of multiple quantum dots for each target antigen.
为克服基于抗体的传感器的局限性，我们描述了一种基于适配体的夹心检测法的概念验证，用于检测与疟疾相关的抗原乳酸脱氢酶。我们展示了在缓冲液中恶性疟原虫乳酸脱氢酶和间日疟原虫乳酸脱氢酶的检测限为0.5飞摩尔，这与基于抗体的检测法相当，使用磁性颗粒 - 适配体构建体进行捕获，以及量子点 - 适配体构建体进行检测。然后，我们使用寡核苷酸功能化的金纳米颗粒，使每个靶抗原能够结合多个量子点，从而证明了检测限为10阿托摩尔（50倍放大）。
