A biomaterial based on a copper complex covalently attached to a polypyrrole backbone was designed formonitoring a glycoprotein, D-dimer, used as a marker of the deep vein thrombosis (DVT) condition. Forthis purpose a new copper complex has been developed based on the ligand N-(2-hydroxybenzyl)-N�-(6-aminohexyl)-N,N�-bis[2-(N-methylimidazolyl)methyl]ethane-1,2-diamine (3) that is able to coordinatecopper ions through two imidazole, two amine and one phenolato moieties—this coordination spherewill be labeled enPI2. The complex conjugated with a polypyrrole layer allows the His-tag antibodyimmobilization onto the conducting polymer substrate and immunosensor evaluation. The biomaterialshows a remarkable variation in redox activity of the Cu(II) complex after the D-dimer interaction. Theredox activity of the [(enPi2)Cu(II)] complex decreases after the antigen interaction providing a linearresponse between 0.01 and 500 ng mL−1with a detection limit of 10 pg mL−1. The chemical structure ofcopper complex demonstrates the ability to avoid non specific-interaction leading to anti fouling surface.Such biolayer architecture offers high measurement stability over time and the biomaterial could be stocked for several weeks without any modification of the electrochemical properties.