The net interaction between a probe tip coated with bovine serum albumin (BSA) protein and a flat substrate coated with poly(ethylene oxide) (PEO) polymer was measured directly on approach in water and electrolyte solutions using AFM. The approach force curve between the two surfaces was monotonically repulsive in water and in electrolyte solutions. At pH ~5, slightly above the isoelectric point (pI) of BSA, and at large distances, the force was dominated by electrostatic repulsion between the oxygen atoms of the incoming protein with those belonging to the ether groups of PEO. Such repulsive force and range decreased in NaCl. Under physiological conditions, pH 6, BSA is definitely charged and the electrostatic repulsion with ether groups in PEO appears at larger separation distances. Interestingly, at pH 4, below the pI of BSA, the repulsion decreased because of an attractive, although weak, electrostatic force that appeared between the ether groups in PEO and the positively charged amino groups of BSA. However, for all solution conditions, once compression of PEO begun, the net repulsion was always dominated by short-range polymeric steric repulsion and repulsive enthalpy penalties for breaking PEO-water bonds. Results suggest that PEO in mushroom conformation may also be effective in reducing biofouling.