Brain tumors are the most common solid tumors in childhood. There is the need for biomarkers of residual disease, response to therapies and recurrence. Cerebrospinal fluid (CSF) is a source of brain tumor biomarkers. We analyzed the proteome of waste CSF from extraventricular drainage (EVD) from 30 children bearing different brain tumors and 16 controls needing EVD insertion for unrelated causes. 1598 and 1526 proteins were identified in CSF control and brain tumor patients, respectively, 263 and 191 proteins being exclusive of either condition. Bioinformatic analysis revealed promising biomarkers for the discrimination between control and tumor (TATA-binding protein-associated factor 15 and S100 protein B). Morever, Thymosin beta-4 (TMSB4X) and CD109, and 14.3.3 and HSP90 alpha could discriminate among other brain tumors and low-grade glyomas plus glyoneuronal tumors/pilocytic astrocytoma, or embryonal tumors/medulloblastoma. Biomarkers were validated by ELISA assay. Our method appears to be able to distinguish among two groups of tumors and each of these from a cohort of both normal and hemorrhagic patients. Further prospective studies may assess whether the biomarkers proposed by our discovery approach can be identified in other bodily fluids, therefore less invasively, and are useful to guide therapy and predict recurrences.