In this work, we investigated the protein adsorption on the end-tethered thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) brushes with varying grafted layer thickness prepared via surface-initiated atom transfer radical polymerization (SI-ATRP) on initiator-immobilized silicon surfaces. The thickness Of a grafted layer was modulated by adjusting reaction time and/or solvent composition. The surface properties as a function of thickness were investigated by water contact angle, X-ray photoelectron spectroscopy (XPS), and atomic force microscope (AFM). The influence of PNIPAAm-grafted layer thickness on human serum albumin (HSA) adsorption in phosphate-buffered saline (PBS) (pH 7.4) at different temperature was evaluated using a radiolabeling method. In a lower thickness range (< 15 nm), protein adsorption on PNIPAAm-grafted layer shows a thermoresponsive change in a certain extent, but the variation is not remarkable. However, it is interesting to observe that these surfaces exhibit good protein-resistant property. For the surface with a PNIPAAm thickness of 13.4 nm, the HSA adsorption level measured at room temperature was similar to 7 ng/cm(2), corresponding to a reduction of 97% compared to the unmodified silicon surface. For thicker PNIPAAm-grafted surface with thickness of 38.1 nm, the adsorption results of three proteins (HSA, fibrinogen, and lysozyme) with different sizes and charges indicate that the PNIPAAm-modified surface exhibits a size-sensitive property of protein adsorption. (C) 2009 Elsevier B.V. All Fights reserved.