Simultaneously Detecting Monoamine Oxidase A and B in Disease Cell/Tissue Samples Using Paper-Based Devices

Abstract

As enzymes in the outer membrane of the mitochondrion, monoamine oxidases (MAOs) can catalyze the oxidative deamination of monoamines in the human body. According to different substrates, MAOs can be divided into MAO-A and MAO-B. The imbalance of the MAO-A is associated with neurological degeneration, while excess MAO-B activity is closely connected with Parkinson’s disease (PD) and Alzheimer’s disease (AD); therefore, detection of MAOs is of great significance for the diagnosis and treatment of these diseases. This work reports the multiplexed detection of MAO-A and MAO-B using paper-based devices based on chemiluminescence (CL). The detection limits were 5.01 pg/mL for MAO-A and 8.50 pg/mL for MAO-B in human serum. In addition, we used paper-based devices to detect MAOs in human cells and tissue samples and found that the results of paper-based detection and Western blotting (WB) showed the same trend. While only one antibody can be incubated on the same membrane by WB, multiple antibodies incubated on the same paper enabled simultaneous detection of MAO-A and MAO-B by paper-based devices. The paper-based assay could be used for preliminary early screening of clinical samples for MAOs and can be extended as an alternative to WB for multiplexed detection of various proteins in disease cell or tissue samples.

Detection of monoamine oxidase B using dark-field light scattering imaging and colorimetry

“…9 In the past few years, various methods have been developed for the quantitative analysis of MAO-B on the basis of enzyme-catalyzed products, such as high-performance liquid chromatography-diode array detection (HPLC-DAD), spectrophotometry, fluorescence, electrochemistry, surface enhanced Raman scattering (SERS) and chemiluminescence (CL). 10–16 However, these traditional methods require a large number of samples or cumbersome detection process. Therefore, developing a label-free and sensitive strategy to monitor MAO-B activity is of great significance for the diagnosis and treatment of neurodegenerative diseases.…”