The effects of hydrophobicity and ligand content on adsorption behaviors of proteins were investigated using dual polarization interferometry (DPI). The silica (DPI chip) surfaces were modified with 3-(aminopropyl) triethoxysilane (APTES), 3-(methacryloxy propyl) trimethoxysilane (MAPTMS) and 3-(diethylaminopropyl) trimethoxysilane (DAPTMS). The content of ligand on DPI chip was controlled by controlling modification time. The adsorption behaviors of bovine serum albumin (BSA), Cytochrome C and chymotrypsin on different surfaces were analyzed. The results indicated that the adsorption capacity of BSA on surface modified with DAPTMS was higher than that modified with APTES and MAPTMS. The adsorption capacity of BSA and adsorption kinetic constant were 1.371 ng/mm2 and 0.056 s?1, respectively. Ligand content affected protein adsorption, which is associated with protein hydrophobicity. BSA and Cytochrome C, as mid-hydrophobic model protein, formed a single layer on DPI surface. The adsorbed capacity of proteins climbs up and then declines with the increase of ligand content. The maximum adsorption capacity was 16.9 nmol/m2 for BSA and 60.2 nmol/m2 for Cytochrome C with N content as 2.1%. Chymotrypsin, as higher hydrophobic model, formed the multi-layer on DPI surface. The adsorption capacity and kinetic constant decrease with the density increases. The max adsorption capacity was 78.6 nmol/m2 and the max kinetic constant was 0.040 s?1 on DPI chip with N content as 1.1%.