Expressed human milk can be donated or sold through a variety of channels, including human milk banks, corporations or individuals, or peer‐to‐peer milk sharing. There is a paucity of research regarding the nutrient and bioactive profiles of expressed human milk exchanged through commerce‐free scenarios, including peer‐to‐peer milk sharing. The study objective was to evaluate the macronutrient, antimicrobial protein, and bacteria composition in expressed human milk acquired via commerce‐free arrangements. Expressed human milk samples were collected from the following commerce‐free scenarios: milk expressed for a mother's or parent's own infant (MOM; N = 30); unpasteurized milk donated to a non‐profit milk bank (BANKED; N = 30); milk expressed for peer‐to‐peer milk sharing (SHARED; N = 31); and health professional‐facilitated milk sharing where donors are serologically screened and milk is dispensed raw (SCREENED; N = 30). Analyses were conducted for total protein, lactose, percent fat and water, lysozyme activity, immunoglobulin A (IgA) activity, total aerobic bacteria, coliform, and Staphylococcus aureus. No bacterial growth was observed in 52/121 samples, and 15/121 had growth greater than 5.0 log colony‐forming units/mL. There was no evidence of differences by groups (p > .05) in lactose, fat, water, lysozyme activity, sIgA activity, aerobic bacteria, coliforms, and S. aureus. Mean protein values (95% confidence interval) were 1.5 g/dL (1.4, 1.6) for BANKED, 1.4 g/dL (1.3, 1.5) for MOM, 1.6 g/dL (1.5, 1.7) for SCREENED, and 1.5 g/dL (1.4, 1.6) for SHARED, which was not significantly different (p = .081). This research contributes to growing literature on the risks and benefits of uncompensated, peer‐to‐peer milk sharing.