The present study reports the achievement of a new chemiluminescent sensing layer able to simultaneously (i) play all active role on ligand immobilization and (ii) serve as a catalyst ill detection processes for label-free biosensor applications. This new type of active Langmuir-Blodgett (LB) monolayer has been designed by a using a chelating lipid (Ni-NTA-DOGS). Thanks to the chelated metallic cation, this peculiar lipid exhibits luminol chemiluminescence catalysis properties in the presence of hydrogen peroxide. Upon biomolecule interaction through imidazole ring chelation (mediated by the metallic cation bound to the lipid headgroup), the chemiluminescent signal call be modulated. The First chemiluminescent signal acquisition experiments have shown a strong and homogeneous signal of the chelating layer. Upon histamine interactions, a histidine derivative used its it market of fresh food quality, we succeeded ill obtaining as it proof of concept a chemiluminescent signal variation without any derivatization of the target molecule. This signal variation was shown to be directly correlated to the histamine concentration with a limit of detection of 2 mu g/mL.