The work aims to study effects of electrophoretic deposition condition and photo-cross linking on morphology and properties of GO/NPs coatings prepared by co-deposition. Firstly, hydroxyethyl methylacrylate (HEMA) was covalently conjugated to the biomacromolecule γ-Polyglutamic acid (γ-PGA) to obtain photo-cross linkable biopolymer γ-PGA-HEMA. Then, γ-PGA-HEMA was used to immobilize the molecular recognition unit horseradish peroxidase (HRP) by electrostatic self-assembly to prepare functional nanoparticles solution. The particle size and morphology of micelles were characterized with nanometer particle size analyzer and TEM. The composite nano-assembly was induced to be co-assembled with GO to prepare multi-component composite deposition solution. The HRP@γ-PGA-HEMA/rGO biological sensing coating was obtained on glass carbon electrode surface by electrophoretic deposition through electrochemical reduction of GO into graphene and control of deposition conditions. Effects of electrophoretic conditions and photo-cross linking on morphology and properties of the coating were investigated by SEM and electrochemical workstation. When electrode position voltage was 1.5 V, electrode position time was 120 sand the GO concentration was 0.1 mg/mL, the coating prepared by electrophoretic deposition exhibited good smoothness, density and uniformity. After photo-cross linking, the compactness and stability of the coating were further improved. The obtained coating showed a high response current to hydrogen peroxide (H2O2). During, the preparation of photo-cross linking coating by electrophoretic deposition composite co-assembly, there is an optimum value for deposition time, deposition voltage and GO content, which endows the electrophoretic coating with complete structure density. The photo-cross linking can further improve the stability of the coating and has the best biosensing performance.