Abstract

Luminescence Approaches for Monitoring Cellular Energy Metabolism

Jolanta Vidugiriene, Senior Scientist, Group Leader, Promega R&D

The growing trend towards understanding the role of cell metabolism in cancer, immunology, obesity, diabetes, and neurodegenerative disease has presented specific challenges in developing rapid and reliable methods for measuring the changes in metabolic pathways. Current technologies are dependent on analytical methods or enzyme couple absorbance/fluorescent assays often require laborious sample preparation and/or lack sensitivity and dynamic range for rapid and convenient metabolite detection directly in 96-well plates. Utilizing novel proluciferin substrate that in the presence of NAD(P)H is converted to luciferin and light production by luciferase, we developed a core technology for measuring key energy co-factors (NAD(P)/NAD(P)H) and extended it to multiple metabolite (glucose, lactate, glutamine, glutamate, glycerol, triglycerides) or enzyme activity assays. Here we demonstrate applicability of those assays to various formats (96-, 384- and 1536-well plates) and samples (2D, 3D cultures) using multiple automation platforms, including collecting the samples at different time points and performing multiple assays for the same sample.