Enhanced Sensitivity of Lateral Flow Strip Biosensors Based on Enyzmatic Reaction and Nanomaterials

• Main Author: Xu, Hui
• Format: Others
• Published: North Dakota State University 2018
• Online Access: https://hdl.handle.net/10365/27289

Ultrasensitive detection for trace amount of proteins plays pivotal role in the diagnosis of specific diseases in clinical application, basic discovery research and the improvement of proteomics. Recently, lateral flow strip biosensor (LFSB) has gained considerable attention for protein analysis. Compared with the traditional immunoassays, LFSB has several advantages: user-friendly format, short assay time (generally several minutes), less interference due to chromatographic separation, a relatively low cost, and no requirements for skilled technicians. This ideal technique is suitable for on-site testing by people who are untrained. Traditional gold nanoparticles (GNPs) based LFSB have been used for qualitative and semiquantitative analysis, the application of GNP-based LFSB is limited by its low sensitivity. In this dissertation, different nanomaterials and advanced detection technologies have been used to enhance the LFSB sensitivities. An ultrasensitive LFSB based on horseradish peroxidase (HRP)/GNP dual labels was developed for qualitative (Yes/No) and quantitative detection of protein. The LFSB signal was enhanced dramatically by introducing the second tracer (enzyme) on the GNP surface. The detection limit of LFSB was 100 times lower than that of GNP-based LFSB. A fluorescent LFSB based on enzyme tracers was developed for sensitive detection of proteins. Alkaline Phosphatase (ALP) was selected as a label to prepare the LFSB. The signal was from the fluorescent emission of the ELF-97 alcohol precipitate which was the product of ALP catalyzed dephosphorylation of ELF-97 phosphate. ALP-conjugated antibody (ALP-Ab) functionalized gold nanoparticles (GNPs) were used as labels for the development of a chemiluminescence-based quantitative LFSB. The use of detection and GNPs as enzyme carriers allowed accurate and sensitive analyte detection. GNP-decorated silica nanorods (GNP-SiNRs) were synthesized and employed as the labels for ultrasensitive detection of proteins on the LFSB. Owing to its biocompatibility and convenient surface modification, SiNRs were used as carriers to load numerous GNPs. The signal of the GNP-SiNR based LFSB was enhanced significantly compared to the GNP-based LFSB since more GNPs were captured through the sandwich-type immunoreactions.