Process for isolating and purifying taste bud receptors for screening and evaluating tastants

摘要

A process for isolating and purifying taste bud receptor material for screening and evaluating tastants is described. The process includes surgically excising a plurality of circumvallate papillae, coring said papillae to remove the central mucous gland from each, followed by exposing the taste bud bearing epithelial portion thereof. The buds are preferably exposed by swelling in a hypotonic medium followed by subjecting said buds to about 900 psi in a nitrogen bomb. The swelled buds are then subjected to a gentle, selective homogenization to extrude taste bud receptor material therefrom without substantial destruction of surrounding epithelial tissue. The extruded material is then isolated by filtering and purified by subjecting the filtrate to differential centrifugation to isolate a receptor containing membrane enriched fraction. The fraction is then further purified by sucrose gradient centrifugation to isolate a receptor containing membrane enriched subfraction substantially without contaminating materials capable of binding tastants. The subfraction is then utilized as a medium for evaluating tastants wherein radioactive labeled tastant compounds are incubated with said subfraction until equilibrium is reached, and the quantity of labeled tastant is bound to the receptor material may then be measured. Unlabeled tastants may also be incubated with said labeled tastant bound medium to evaluate the strength of the binding, and other related taste characteristics.