PurposeSweat is a relatively unexplored biofluid for diagnosis and monitoring of disease states. In this study, the proteomic profiling of immune‐related biomarkers from healthy individuals are presented.Experimental DesignEccrine sweat samples are collected from 50 healthy individuals. LC‐MS/MS is performed on two pools of sweat samples from five male and female participants. Individual sweat samples are analyzed by antibody isotyping microarrays (n = 49), human cytokine arrays (n = 30), and quantitative ELISAs for interleukin‐1α (n = 16), epidermal growth factor (n = 6), and angiogenin (n = 7).ResultsIn sweat, 220 unique proteins are identified by shotgun analysis. Detectable antibody isotypes include IgA (100% positive; median 1230 ± 28 700 pg mL−1), IgD (18%; 22.0 ± 119 pg mL−1), IgG1 (96%; 1640 ± 6750 pg mL−1), IgG2 (37%; 292 ± 6810 pg mL−1), IgG3 (71%; 74.0 ± 119 pg mL−1), IgG4 (69%; 43.0 ± 42.0 pg mL−1), and IgM (41%; 69.0 ± 1630 pg mL−1). Of 42 cytokines, three are readily detected in all sweat samples (p < 0.01). The median concentration for interleukin‐1α is 352 ± 521 pg mL−1, epidermal growth factor is 86.5 ± 147 pg mL−1, and angiogenin is 38.3 ± 96.3 pg mL−1. Multiple other cytokines are detected at lower levels.Conclusions and Clinical RelevanceSweat can be used for profiling antibodies and innate immune biomarkers.