Niemann Pick Type C (NPC) Disease is a neurodegenerative disease that almost invariably leads to death in pediatric patients. There is currently no known cure for the disease. Intracellular accumulation of cholesterol and other lipids occur in NPC and in other related neurodegenerative diseases. Theta toxin produced by bacteria binds to cholesterol with high affinity. Biotinylated theta toxin (BC-theta) has been shown to be an effective cholesterol stain. Purified theta toxin is not commercially available. In this work we developed a simple method to isolate and purify the theta toxin, by using lysates of bacterial cells that express the recombinant histidine-tagged theta toxin as the starting material. The improved method consists of running a single nickel affinity column chromatography followed by dialysis and biotinylation of the purified toxin protein. The improved method will prove useful in future cell imaging studies.

Improved procedure for purifying the theta toxin suitable for imaging intracellular cholesterol accumulation in mammalian cells

244th National Meeting (2012)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.