As a selective prebiotic, 2'-FL significantly promotes the proliferation of beneficial bacteria such as Bifidobacterium, while inhibiting the growth of harmful bacteria like Clostridioides difficile, helping to establish a healthy gut microecological environment. Clinical studies have shown that supplementation with 2'-FL in infant formula can increase the relative abundance of Bifidobacterium in the infant gut to approximately 60%, approaching the levels observed in breastfed infants. Fermentation of 2'-FL by gut microbiota enhances the production of short-chain fatty acids (acetate and butyrate), supporting anti-inflammatory responses and providing energy for colonocytes.

2'-FL promotes immune system maturation, elevates plasma IgG and IgA levels, and strengthens the intestinal mucosal barrier. A randomized controlled clinical trial involving over 300 infants demonstrated that those fed formula supplemented with 2'-FL exhibited 29%–83% lower concentrations of inflammatory cytokines (IL-1β, IL-6, TNF-α). Another clinical study showed that formula supplemented with 2'-FL and LNnT reduced the risk of upper respiratory tract infections by 42% and otitis media by 71%, while also significantly reducing the use of associated medications.

2'-FL acts as a "decoy receptor" that binds to harmful pathogens, preventing their attachment to intestinal epithelial cells and helping to flush them out of the body. Studies have shown that 2'-FL can reduce Escherichia coli O157 intestinal colonization by over 90% and Campylobacter jejuni invasion by 80%, while significantly enhancing mucin MUC2 expression (by more than 20%), strengthening the intestinal barrier function. 2'-FL also significantly increased the abundance of Akkermansia, an important beneficial bacterium closely associated with metabolic health.

Research indicates that 2'-FL has positive effects on learning and memory. Animal studies found that pigs fed 2'-FL showed larger olfactory bulb volumes, with altered expression levels of genes related to dopamine, GABA, and choline synthesis. Long-term oral administration of 2'-FL has been shown to improve synaptic plasticity in rodents and increase the expression of molecules involved in storing newly acquired memories, thereby improving learning and memory abilities. Early exposure to 2'-FL may promote infant cognitive development.

Clinical research has shown that infants fed formula supplemented with 2'-FL had colic incidence rates comparable to the breastfed group (10.5% vs 8.8%), and atopic dermatitis incidence was also lower than in the standard formula group. These findings suggest that 2'-FL supplementation may help reduce the risk of infantile colic and atopic dermatitis.

Animal studies have demonstrated that 2'-FL can not only increase insulin levels but also improve the digestibility and utilization of calcium, phosphorus, crude fat, and total energy, contributing to bone health and overall nutrient absorption.