Autoantibodies against tumor-associated antigens in sputum as biomarkers for lung cancer

•It is important to develop biomarkers for the early detection of lung cancer.•By probing a protein microarray containing more than 9000 antigens with sputum of lung cancer patients and cancer-free smokers, we identified 28 lung tumor antigens (TAs).•A panel of three TA-associated autoantibody (TAAb) biomarkers was further developed that had 81% sensitivity and 83% specificity for diagnosis of lung cancer.•This study provides the first evidence for the potential of sputum TAAbs as biomarkers for the early detection of lung cancer. Tumor antigens (TAs) can initiate host immune responses and produce TA-associated autoantibody (TAAbs), potential cancer biomarkers. Sputum is directly generated from the upper and lower airways, and thus can be used as a surrogate sample for the diagnosis of lung cancer based on molecular analysis. To develop sputum TAAb biomarkers for the early detection of lung cancer, the leading cause of cancer death, we probed a protein microarray containing more than 9,000 antigens with sputum supernatants of a discovery set of 30 lung cancer patients and 30 cancer-free smokers. Twenty-eight TAs with higher reactivity in sputum of lung cancer cases vs. controls were identified. The diagnostic significance of TAAbs against the TAs was determined by enzyme-linked immunosorbent assays (ELISAs) in sputum of the discovery set and additional 166 lung cancer patients and 213 cancer-free smokers (validation set). Three sputum TAAbs against DDX6, ENO1, and 14–3-3ζ were developed as a biomarker panel with 81% sensitivity and 83% specificity for diagnosis of lung cancer, regardless of stages, locations, and histological types of lung tumors. This study provides the first evidence that sputum TAAbs could be used as biomarkers for the early detection of lung cancer.