Human Phospholipase A₂ (PLA₂) has been identified as a potential biomarker for atherosclerosis, cancer, sepsis, and other inflammatory conditions. PLA₂ catalyzes the hydrolysis of phospholipids. Lipoprotein-associated PLA₂ (Lp-PLA₂), an isoform of human PLA₂, has emerged as a selective biomarker for early diagnosis of cardiovascular disease (CVD). Even though several CVD screening and diagnostic methods are available, they have inherited limitations that prevent selective and sensitive early diagnosis of CVD. Currently, enzyme linked immunosorbent assay (ELISA) is considered the state-of-the-art technique in detecting Lp-PLA₂. In ELISA a two-step sandwich immunoassay is utilized to target antibodies to the Lp-PLA₂ to measure either the enzyme concentration or the activity. Though ELISA-based techniques are generally sensitive, they have inherent drawbacks for early diagnosis that must be addressed by developing new approaches. In this study, we will present the development of a microfluidic-based detection method for PLA₂. A simple microfluidic device with supported lipid membranes (SLMs) is constructed using soft lithographic techniques The SLM consists of a mixture of lipids, including anionic, zwitterionic, and fluorescent lipids, which are reactive with PLA₂. The decrease in fluorescence due to the disintegration of SLMs upon hydrolysis is monitored using fluorescence microscopy followed by ImageJ analysis. In this presentation, we will discuss our success in the detection of PLA₂ via microfluidic approaches.

Menake Piyasena

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