Immunological Detection of in Vitro Formed Phosphatidylethanol—An Alcohol Biomarker—With Monoclonal AntibodiesAlcoholism: Clinical and Experimental Research OnlineEarly Articles 28 Apr 2008
Phosphatidylethanol (PEth) is a promising new marker for detecting long-term alcohol abuse with excellent sensitivity and specificity. Current methods are based on the high performance liquid chromatography–mass spectrometric method and therefore require high levels of expertise and expensive instrumentation.
This study was designed to generate PEth-specific monoclonal antibodies for PEth immunoassay development.
We generated monoclonal cell lines (2B1 and 2E9) that produce IgM antibodies binding specifically to PEth but not to structurally or chemically similar phospholipids, such as phosphatidylcholine, phosphatidic acid, and cardiolipin. We show here that these anti-PEth antibodies can be used to detect PEth in a fluorescent PEth assay and FACS analysis of human red blood cell samples spiked with PEth.
The present study shows that PEth-specific monoclonal antibodies can be generated using traditional hybridoma technique. Immunogenicity of PEth was enhanced using human LDL as an immunization carrier. The generated monoclonal anti-PEth antibodies, 2B1 and 2E9 bind to PEth in fluid phase and in biological membranes.
Request Reprint E-Mail: markku.savolainen@oulu.fi
_________________________________________________________________
