Bronchoalveolar lavage fluid (BALF) is a biological fluid that is comprised of airway epithelial, immune cells, and soluble materials. It reflects the cellular and molecular states of the lung and can provide critical insights into pathogenesis of pulmonary diseases as well as valuable biomarker candidates. However, the BALF proteome is dominated by plasma-derived high-abundance proteins that lead to a large dynamic range creating a challenge for comprehensive profiling. Herein we address this challenge by developing a rapid four-dimension LC-MS approach using trapped ion mobility spectrometry (TIMS) and parallel accumulation serial fragmentation (PASEF) with a data-independent acquisition (DIA) method. Our workflow facilitates high-throughput capability that achieves deep proteome coverage and high quantitative reproducibility for BALF analysis.