BackgroundIdiopathic Normal pressure hydrocephalus (iNPH) is a form of adult hydrocephalus that is clinically characterized by progressive gait impairment, cognitive dysfunction, and urinary incontinence. The current standard method of treatment involves surgical installation of a CSF diversion shunt. However, only a fraction of patients shows an alleviation of symptoms from shunt surgery. Thus, the purpose of this prospective explorative proteomic study was to identify prognostic CSF biomarkers to predict shunt responsiveness in iNPH patients. Further, we evaluated the ability of the core Alzheimer’s disease (AD) CSF biomarkers phosphorylated (p)-tau, total (t)-tau, and amyloid-β 1–42 (Aβ1–42) to serve as predictors of shunt response.MethodsWe conducted a tandem mass tag (TMT) proteomic analysis of lumbar CSF from 68 iNPH patients, sampled pre-shunt surgery. Tryptic digests of CSF samples were labelled with TMTpro reagents. The TMT multiplex samples were fractionated in 24 concatenated fractions by reversed-phase chromatography at basic pH and analysed by liquid chromatography coupled to mass spectrometry (LC–MS) on an Orbitrap Lumos mass spectrometer. The relative abundances of the identified proteins were correlated with (i) iNPH grading scale (iNPHGS) and (ii) gait speed change 1 year after surgery from baseline to identify predictors of shunt responsiveness.ResultsWe identified four CSF biomarker candidates which correlated most strongly with clinical improvement on the iNPHGS and were significantly changed in shunt-responsive compared to shunt-unresponsive iNPH patients 1 year post-surgery: FABP3 (R= − 0.46, log2(fold change (FC)) = − 0.25,p< 0.001), ANXA4 (R= 0.46, log2(FC) = 0.32,p< 0.001), MIF (R= -0.49, log2(FC) =  − 0.20,p< 0.001) and B3GAT2 (R= 0.54, log2(FC) = 0.20,p< 0.001). In addition, five biomarker candidates were selected based on their strong correlation with gait speed change 1 year after shunt installation: ITGB1 (R= − 0.48,p< 0.001), YWHAG (R= − 0.41,p< 0.01), OLFM2 (R= 0.39,p< 0.01), TGFBI (R= − 0.38,p< 0.01), and DSG2 (R= 0.37,p< 0.01).Concentrations of the CSF AD core biomarkers did not differ significantly with shunt responsiveness.ConclusionFABP3, MIF, ANXA4, B3GAT2, ITGB1, YWHAG, OLFM2, TGFBI and DSG2 in CSF are promising prognostic biomarker candidates to predict shunt responsiveness in iNPH patients.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12987-023-00440-5.