Chronic obstructive pulmonary disease (COPD) is the second leading cause of death in India, primarily attributed to smoking. Asymptomatic smokers can develop COPD due to a combination of genetic, environmental, and molecular factors, highlighting the importance of early detection. Data-independent acquisition mass spectrometry (DIA-MS) proteomics provides an unbiased approach to analyzing proteomic profiles. This study is the first to utilize DIA-based proteomics to examine individual serum samples from three distinct male groups: healthy individuals (n=10), asymptomatic smokers (n=10), and COPD patients (n=10). This thorough analysis identified 667 proteins with a 1% false discovery rate. Differential expression analysis revealed 40 proteins distinguishing normal from asymptomatic smokers, 88 proteins differentiating COPD from healthy individuals, and 40 proteins differentiating COPD from asymptomatic smokers. Notably, proteins such as PRDX6, ELANE, PRKCSH, PRTN3, and MNDA may help distinguish COPD from asymptomatic smokers, while ELANE, H3-3A, IGHE, SLC4A1, and SERPINA11 could differentiate COPD from healthy subjects. Pathway enrichment and protein-protein interaction analyses highlighted significant changes in hemostasis, immune system functions, fibrin clot formation, and post-translational protein modifications. Key proteins were validated through a parallel reaction monitoring assay. Our findings identify critical protein markers in COPD patients, asymptomatic smokers, and healthy individuals, providing insights for clinicians to better understand the disease’s pathobiology, enhance disease management, and improve patient quality of life