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The use of <i>Opuntia ficus-indica</i> fruits in the agro-food sector is increasing for a multiplicity of players. This renewed interest is, in part, due to its organoleptic characteristics, nutritional value and health benefits. Furthermore, industries from different sectors intend to make use of its vast array of metabolites to be used in different fields. This trend represents an economic growth opportunity for several partners who could find new opportunities exploring non-conventional fruits, and such is the case for <i>Opuntia ficus-indica</i>. <i>O. ficus-indica</i> originates from Mexico, belongs to the Cactaceae family and is commonly known as opuntia, prickly pear or cactus pear. The species produces flowers, cladodes and fruits that are consumed either in raw or in processed products. Recent publications described that consumption of the fruit improves human health, exhibiting antioxidant activity and other relevant pharmacological activities through enzymatic and non-enzymatic mechanisms. Thus, we provide a systematic, scientific and rational review for researchers, consumers and other relevant stakeholders regarding the chemical composition and biological activities of <i>O. ficus-indica</i> fruits.