This project encompasses the serum proteomic data generated for the discovery of novel biomarkers for colorectal cancer (CRC). Utilizing data-independent acquisition (DIA) mass spectrometry, we comprehensively profiled the proteomes of serum samples from individuals with CRC, colorectal adenoma, and healthy controls. The analysis identified a panel of significantly dysregulated proteins, highlighting altered pathways in CRC pathogenesis. From this, ROBO4 and VCAM-1 were selected as top candidate biomarkers based on their significant differential expression and relevance to cancer biology. This dataset provides a valuable resource for exploring the serum protein landscape in colorectal carcinogenesis and for the discovery of diagnostic signatures.