The establishment and public sharing of comprehensive proteome maps of human tissues and biofluids are essential to improve biomedical knowledge and to support biomarker discovery studies. In this study, we performed an in-depth proteomics characterization of the exhaled breath condensate (EBC), a non-invasive sample obtained from the condensation of the airway lining fluid. Two pooled samples of EBC, each combined from 10 healthy donors, were lyophilized and their protein content analyzed by high-resolution tandem mass spectrometry-based proteomics. Two “technical” negative control samples were processed in parallel with the pooled samples to correct for exogenous protein contamination and avoid erroneous identifications.