Vanishing white matter (VWM) disease is a genetic leukodystrophy leading to severe neurological disease and early death. VWM is caused by bi-allelic mutations in any of the five genes encoding the subunits of the eukaryotic translation factor 2B (EIF2B). Although previous studies are being attempted to investigate the molecular mechanism of VWN by constructing variant models for each subunit of EIF2B that causes VWM disease. The underlying molecular mechanism of how mutations (MT) in EIF2B3 result in VWM is unknown. Based on our recently report, we constructed an eif2b3 knockout (eif2b3-/-) zebrafish (Danio rerio) model and performed quantitative proteomic analysis between the wide type (WT) and eif2b3-/- models to identify 25 differentially expressed proteins (DEPs). A total of 4 proteins were significantly up-regulated, and 21 proteins were significantly down-regulated in eif2b3-/- larvae compared to WT, respectively. Representatively, Lon protease and neutral amino acid transporter SLC1A4 were significantly increased in truncated eif2b3 zebrafish, and crystallin proteins were also significantly decreased. In conclusion, this study proposed the candidate proteins as a key regulator related to the progression of VWN disease through quantitative proteomic analysis in the first variant model of VWN disease.