2'-Fucosyllactose (2'-FL) is a trisaccharide composed of fucose, galactose, and glucose. It is the most abundant human milk oligosaccharide (HMO) found in breast milk. Its chemical structure features a fucose residue linked via an α1-2 glycosidic bond to the galactose unit of lactose. The primary application of 2'-FL is as a key functional ingredient in infant formula to mimic the composition and benefits of human milk. It acts as a prebiotic, selectively promoting the growth of beneficial gut bacteria like *Bifidobacteria*. Furthermore, it plays a crucial role in supporting immune system development, protecting against pathogens by acting as a decoy receptor, and supporting brain development in infants. Due to these significant health benefits, it is a high-value additive in the nutritional and pediatric health sectors. Beyond infant nutrition, 2'-FL is also being investigated for its potential applications in adult nutritional products, functional foods, and dietary supplements aimed at gut health and immune modulation. Its production through advanced biotechnological fermentation has made commercial-scale supply feasible.