Cholesterol esters are synthesized in plasma from cholesterol and an acyl chain on phosphatidylcholine via a reaction catalyzed by lecithin:cholesterol acyltransferase (LCAT). Another mechanism for making cholesterol esters is via the enzymatic reaction catalyzed by ACAT.

Cholesterol esters are considerably more hydrophobic than cholesterol itself. The amounts of cholesterol and cholesterol esters associated in blood lipoprotein complexes called LDL are typically about two-thirds of the total plasma cholesterol (total plasma cholesterol ranges from 130 to 260 mg/100 mL of human plasma, with the most desirable levels between 160 and 200). More than 40% of the weight of the LDL particle is cholesterol esters, and the total of esterified and free cholesterol amounts to well over half the total weight.