Update publication information. Anlotinib has been demonstrated to be effective in advanced non-small cell lung cancer (NSCLC) patients. DNA-seq- and RNA-seq-based anlotinib molecular mechanism and biomarker screening have been introduced in previous studies. The underlying value of proteomics for anlotinib study is still unclear. In this study, 70 plasma samples were selected from 28 anlotinib-treated NSCLC patients (including 14 responders and 14 non-responders). LC-MS/MS analysis was performed on those samples with different time points including baseline, best response and progression disease. Bioinformatics analysis was performed to understand the underlying value of those differential proteins. Proteomics analysis suggested the differential proteins from responders after anlotinib administration potential play a role in the molecular mechanism characterization and biomarker screening. The differential proteins between responders and non-responders at baseline mainly contribute to biomarker screening. Integrative analysis indicated 43 proteins could be used as underlying biomarkers for clinical practice. Lastly, we selected ARHGDIB and demonstrated that it has some predictive value for anlotinib. Collectively, this study not only offered the first insight that the proteomic technology potentially be used for anlotinib molecular mechanism characterization, but also provided a basis for anlotinb biomarker screening via proteomics in the future.