核酶介导的质量敏感生物传感器信号增强

Ribozyme-mediated signal augmentation on a mass-sensitive biosensor.

作者信息

Knudsen Scott M, Lee Joonhyung, Ellington Andrew D, Savran Cagri A

机构信息

Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas 78712, USA.

出版信息

J Am Chem Soc. 2006 Dec 20;128(50):15936-7. doi: 10.1021/ja064137m.

DOI:10.1021/ja064137m

PMID:17165697

Abstract

Mass-based detection methods such as the quartz crystal microbalance (QCM) offer an attractive option to label-based methods; however the sensitivity is generally lower by comparison. In particular, low-molecular-weight analytes can be difficult to detect based on mass addition alone. In this communication, we present the use of effector-dependent ribozymes (aptazymes) as reagents for augmenting small ligand detection on a mass-sensitive device. Two distinct aptazymes were chosen: an L1-ligase-based aptazyme (L1-Rev), which is activated by a small peptide (MW approximately 2.4 kDa) from the HIV-1 Rev protein, and a hammerhead cleavase-based aptazyme (HH-theo3) activated by theophylline (MW = 180 Da). Aptazyme activity was observed in real time, and low-molecular-weight analyte detection has been successfully demonstrated with both aptazymes.

摘要

基于质量的检测方法，如石英晶体微天平（QCM），为基于标记的方法提供了一个有吸引力的选择；然而，相比之下，其灵敏度通常较低。特别是，仅基于质量增加来检测低分子量分析物可能会很困难。在本通讯中，我们展示了使用效应物依赖性核酶（适体酶）作为试剂，以增强在质量敏感设备上对小配体的检测。选择了两种不同的适体酶：一种基于L1连接酶的适体酶（L1-Rev），它由来自HIV-1 Rev蛋白的一种小肽（分子量约2.4 kDa）激活；另一种基于锤头状切割酶的适体酶（HH-theo3），由茶碱（分子量 = 180 Da）激活。实时观察到了适体酶活性，并且两种适体酶都已成功证明了对低分子量分析物的检测。