Multiple sclerosis (MS) is a chronic autoimmune, inflammatory neurological disease of the central nervous system (CNS) resulting in damage to myelin and myelinated axons in the CNS and can lead to severe disability. The most common form, relapsing-remitting MS (RRMS), is marked by acute onset of neurologic symptoms (relapses), followed by periods of remission. Another form, primary progressive MS (PPMS) is a severe manifestation of the disease characterized by unabated worsening symptoms without remission. In order to identify candidate biomarkers of disease activity, or for use in evaluation of therapeutic response across different manifestations of the disease, an in-depth proteome analysis of CSF from two independent cohorts of RRMS, PPMS, and control patients with both healthy and other neurologic diseases (OND) was conducted. Mass spectrometry-based proteomics was employed using a data independent acquisition (DIA) workflow with a CSF-specific spectral library. Results from two independent cohorts were compared and proteins with differential CSF expression patterns across disease subtypes were identified for further characterization. This DIA approach allowed for the identification of greater than 1,500 proteins quantified across the samples. Statistical testing allowed for the identification of protein groups significantly changed in PPMS, RRMS or PPMS samples compared to RRMS samples. Pathway analysis revealed changes between RRMS and PPMS in pathways that included B cell and macrophage biology. This extensive study gives a comprehensive analysis of the RRMS and PPMS proteome observed from CSF that will be used to inform clinical biomarker strategies focused on evaluating therapeutic mechanism of action. Herein we provide a comprehensive profile of the MS CSF proteome and identify biomarker candidates for further investigation.