Human 2-arachidonoylglycerol (2-AG) is an agonist of endocannabinoid system and acts as an important modulator of many physiological processes such as emotional state and pain sensation. Identification and quantification of 2-AG is vital for medical and pathological processes. There are no reports on the measurement of 2-AG in human biofluids using modern methods such as biosensors. This study reports an ultra-sensitive and selective immunosensor to determine endocannabinoids 2-AG in human plasma samples. In this study, gold nano-flowers (AuNFs) were synthesized and conjugated with a specific biotinylated antibody of 2-AG. Bioconjugated composite (bioreceptor with AuNFs) was immobilized on the surface of a gold electrode and used for the monitoring of the antigen (target molecules) based on the immunoreaction process. Moreover, a constructed interface was characterized by field-emission scanning electron microscopy (FE-SEM), dynamic light scattering (DLS), transmission electron microscopy (TEM) and zeta potential methods. Using the proposed immuno-platform, 2-AG was determined in two dynamic ranges of 0.00024-0.0078 ng L-1 and 2-16 ng L-1 with a lower limit of quantitation (LLOQ) of 0.00024 ng L-1. These results suggest that our immunosensor might be appropriate for an early diagnosis of 2-AG towards the screening of immunomodulatory activity and neuroprotection.