Considerable research has been focused on the exploration of bone morphogenetic protein-2 (BMP₂) delivery vehicles for achieving prolonged availability and maintaining efficient local concentration at the bone injury sites. In this study, heterobifunctional cross-linkers Sulfo-SMCC and cyclic thioimidate compound Traut's Reagent were used to conjugate monoclonal polyhistidine antibody on collagen scaffold demineralized bone matrix (DBM) to create specific binding between BMP₂ containing six histidines tag (His-BMP₂) and DBM. According to the optimized cross-linking reagent concentration, more polyhistidine antibodies conjugated on DBM with 5mg/ml Traut's Reagent and 25ug/ml Sulfo-SMCC than physical adsorption. Monoclonal antibodies conjugated DBM (MAbs-DBM) could bind more His-BMP₂ than DBM and achieved controlled release in vitro. The alkaline phophatase (AP) activity of C2C12 cells on MAbs-DBM indicated that His-BMP₂ retained on MAbs-DBM preserved the function to induce osteogenic differentiation. His-BMP₂/MAbs-DBM induced more ectopic bone formation (AP activity assay and histochemistry stain) than control group after subcutaneous implantation. The results demonstrated that antibody-collagen system could be useful for maintaining higher local therapy concentration of growth factors at the injury sites.