Abstract
Proteases play a pivotal role in regulating important physiological processes from food digestion to blood clotting. They are also important biomarkers for many diseases such as cancers. The importance of proteases has led to extensive efforts in the screening of proteases and their inhibitors as potential drug molecules. For example, human immunodeficiency virus (HIV) patients have been treated with HIV-1 protease inhibitors to prolong the life expectancy of patients. Such a close relationship between diseases and proteases provides a strong motivation for developing sensitive, selective, and robust protease assays and sensors, which can be exploited to discover new proteases and inhibitors. In this aspect, protease assays based on levels of proteolytic activities are more relevant than protease affinity assays such as immunoassays. In this review, recent developments of protease activity assays based on different detection principles are discussed and compared. For homogenous assays, fluorescence-based techniques are the most popular due to their high sensitivity and quantitative results. However, homogeneous assays have limited multiplex sensing capabilities. In contrast, heterogeneous assays can be employed to detect multiple proteases simultaneously, given the microarray technology that is already available. Among them, electrochemical methods, surface spectroscopy techniques, and enzyme-linked peptide protease assays are commonly used. Finally, recent developments in liquid crystal (LC)-based protease assays and their applications for detecting proteases and their inhibitors are discussed.
Highlights
Proteases are enzymes that catalyse the hydrolysis of peptide bonds in peptide chains
Analyst influence anchoring of liquid crystal (LC) by engineering the chemical composition or morphology of surfaces supporting the LC phase.[89,90]. This phenomenon was rigorously studied by Abbott and coworkers, who established principles to detect the adsorption of biomolecules on solid surfaces or LC/water interfaces.[91,92,93,94,95,96,97,98,99]
Branchini et al incorporated an organic fluorophore in their assay, where a red fluorescent protein (RFP) was labelled with Alexa Fluor® (AF) 680 dye and linked to a luciferase (Luc) through a specific peptide substrate.[126]
Summary
Proteases play a pivotal role in regulating important physiological processes from food digestion to blood clotting. Human immunodeficiency virus (HIV) patients have been treated with HIV-1 protease inhibitors to prolong the life expectancy of patients Such a close relationship between diseases and proteases provides a strong motivation for developing sensitive, selective, and robust protease assays and sensors, which can be exploited to discover new proteases and inhibitors. In this aspect, protease assays based on levels of proteolytic activities are more relevant than protease affinity assays such as immunoassays. Recent developments in liquid crystal (LC)-based protease assays and their applications for detecting proteases and their inhibitors are discussed
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