Detection of glycolipids as biotinylated derivatives using enhanced chemiluminescence.

A new method for the detection of glycolipids as biotinylated derivatives is presented. This method is based on the detection of lipids by enhanced chemiluminescence (ECL) following conjugation with streptavidin-horseradish peroxidase (SHRP). Partial biotinylation of glycolipids is achieved after mild oxidation of the glycan moiety with Na-meta-periodate to increase the availability of biotin-reactive sites. There are several significant advantages of the glycolipid ECL detection: it avoids the need for radio-labeling; it provides the possibility of antibody probing; it allows reprobing of SHRP conjugate to adjust background and luminol light emission levels; it permits easy and accurate quantification of glycolipids at low concentrations; and it involves nondestructive staining, thereby enabling further molecular analysis.