The neuroprotective BDNF/TrkB-FL system is compromised in Alzheimer’s disease (AD). Amyloid-beta triggers calpain-mediated TrkB-FL receptor cleavage, leading to TrkB-ICD formation, a novel intracellular toxic fragment. Biological fluids are used to pinpoint potential disease biomarkers and extracellular vesicles (EVs) are cell-specific carriers of promising pathological biomarkers. Thus, this work aimed to investigate the presence of TrkB-ICD in AD human samples and EVs. Protein and RNA was extracted from human AD post-mortem brain samples classified according to the Braak staging (stages 0-II, III-IV). For CSF and plasma samples, patients fulfilled the criteria for Mild Cognitive-Impairment due to AD – high likelihood (MCIAD), whereas controls (MCICONTROL) reported cognitive complaints but had no evidence of Aβ pathology or neuronal injury. MCIAD plasma-derived EVs (pdEVs) were isolated using the ExoQuick reagent and characterized. EVs from 48-hour conditioned medium of control, GFP- and TrkB-ICD-V5-transduced differentiated SH-SY5Y cells were also isolated through differential ultracentrifugation and characterized to better understand TrkB-ICD EV incorporation. Human post-mortem samples revealed a decrease in TrkB-FL protein levels concomitantly with an increase in TrkB-ICD fragment levels (p=7.3 x10-3 and 3.9 x10-3, n=7-11) with AD progression. CSF analysis showed increased TrkB-ICD immunoreactivity in MCIAD when compared to MCICONTROL patients (p=7.55x10-3, n=23-47), and a negative correlation between the levels of Aβ1-42 and TrkB-ICD (ρ=-0.47, n=47). Regarding EV presence, plasma of MCIAD patients contained higher levels of TrkB-ICD (p=0.010, n=17-18). TrkB-ICD and TrkB-ICD-V5 were detected in both SH-SY5Y EV subpopulations equally (p>0.05, n=3). Altogether, these data demonstrate TrkB-ICD extracellular secretion, alluding for its potential toxicity dissemination.