Apolipoprotein A-I (ApoA-I) is the primary protein component of high density lipoproteins (HDL). ApoA-I plays an important role in cholesterol metabolism by mediating the formation of nascent HDL and the efflux of cellular cholesterol from macrophage foam cells in arterial walls. Lipidation of ApoA-I is mediated by the ATP-binding cassette A1 (ABCA1). Insufficient ABCA1 activity my lead to reduced HDL formation, reduced cholesterol efflux and the development of arteriosclerosis.The standard radioactive assay for measuring cholesterol transport to lipid-free ApoA-I has low through-put, poor dynamic range and fails to measure phospholipid transferred with cholesterol. We describe the development of two sensitive, non-radioactive high-throughput assays that report on the lipidation state of ApoA-I and may have applications for studying ABCA1 function and HDL metabolism: a homogenous assay based on the Time Resolved FRET (TR-FRET) and a discontinuous assay that uses the Epic.The TR-FRET assay employs a fluorescent ApoA-I where Cysteine is labeled with the FRET acceptor HiLyte-Fluor-647 and an N-terminal Biotin-AviTag is bound to the streptavidin-Terbium conjugate. When this ApoA-I was incorporated into recombinant HDL, TR-FRET decreased proportionally to the increase in the ratio of lipid to ApoA-I in agreement with the expansion of the surface area of lipids concomitant with the increase in separation of N-terminal and central regions of the protein and demonstrated that the HTRF assay was sensitive to the amount of lipid associated with ApoA-I.The Epic is a label-free platform that allows for the observation of direct biomolecular interactions via a resonant wavelength shift which is proportional to the mass bound to the surface. In the Epic assay, biotinylated ApoA-I was captured on streptavidin-coated sensor. The response was proportional to the amount of lipids associated with ApoA-I indicating that the assay could sense lipidation of ApoA-I.