Chen Yu-Hsuan, Gupta Nitesh K, Huang Hsiao-Jung, Lam Chak Hin, Huang Ching-Lan, Tan Kui-Thong
Department of Chemistry, National Tsing Hua University, 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University, 101 Section 2, Kuang Fu Road, Hsinchu 30013, Taiwan, Republic of China.
Anal Chem. 2021 Apr 6;93(13):5556-5561. doi: 10.1021/acs.analchem.1c00138. Epub 2021 Mar 25.
Lateral flow assay (LFA) has been a valuable diagnostic tool in many important fields where rapid, simple, and on-site detection is required, for applications such as pregnancy tests and infectious disease prevention. Currently, two types of LFAs are available: lateral flow immunoassay (LFIA) and nucleic acid lateral flow assay (NALFA). Both are generally used for the testing of proteins and nucleic acids. However, enzyme activities and small molecules without the corresponding binding partner cannot be detected by the existing LFAs. In this paper, we introduce a LFA approach termed ffinity-witchable ateral low ssay (ASLFA) to overcome the limitations. The detection principle is based on the switchable binding between the affinity-switchable biotin (ASB) probe and avidin protein. In the presence of the target molecule, the activated ASB probe would be captured by the avidin, thereby leaving a distinct test line on the membrane. The ASLFA concept was demonstrated by testing the F ion, NADH cofactor, and nitroreductase activity. Thus, this general ASLFA can be used for the rapid detection of molecules that cannot be accessed by the classical LFAs.
侧向流动分析(LFA)在许多需要快速、简单和现场检测的重要领域中一直是一种有价值的诊断工具,用于诸如妊娠测试和传染病预防等应用。目前,有两种类型的LFA:侧向流动免疫分析(LFIA)和核酸侧向流动分析(NALFA)。两者通常都用于蛋白质和核酸的检测。然而,现有的LFA无法检测酶活性和没有相应结合伴侣的小分子。在本文中,我们引入了一种称为亲和可切换侧向流动分析(ASLFA)的LFA方法来克服这些局限性。检测原理基于亲和可切换生物素(ASB)探针与抗生物素蛋白之间的可切换结合。在存在目标分子的情况下,活化的ASB探针将被抗生物素蛋白捕获,从而在膜上留下明显的测试线。通过检测F离子、NADH辅因子和硝基还原酶活性证明了ASLFA的概念。因此,这种通用的ASLFA可用于快速检测经典LFA无法检测的分子。