Severe traumatic brain injury (sTBI) has become a global health issue and has caused significant social and family burdens worldwide. Although proteomic techniques have discovered some biomarkers of sTBI, the common biomarkers and pathological mechanisms of sTBI at different time points have not been systematically revealed. In recent years, studies have found significant changes in the expression of ubiquitin and ubiquitination modified proteins after TBI, but the specific ubiquitination modified proteins that undergo changes are still unclear. This study used label free ubiquitination proteomics technology to detect and screen for common differential ubiquitination modification proteins and sites in the sTBI group at different time points compared to the control group. Bioinformatics analysis was used to explore the pathological mechanism mediated by ubiquitination modification proteins after the occurrence of sTBI.Finally, parallel reaction monitoring (PRM) validation experiments were conducted on the ubiquitination sites of proteins in the selected specimens, confirming the omics resultsproviding new diagnostic and therapeutic ideas for the treatment of sTBI.