Enzymatic hydrolysis of lignocellulosic materials is significantly affected by the cellulase adsorption onto the cellulosic substrates and lignin. The presence of lignin plays an important role in the lignocellulosic hydrolysis and enzyme recycling. Three cellulase preparations (Celluclast, Spezyme CP and MSUBC) were evaluated to determine their adsorption onto cellulolytic enzyme lignin (CEL) from steam exploded Lodgepole line (SELP) and ethanol (organosolv) pretreated Lodgepole pine (EPLP). The adsorption affinity of cellulase (Celluclast) onto isolated lignin (CEL-EPLP and CEL-SELP) was slightly higher than that from corresponding EPLP and SELP substrates based on the Langmuir constants. Effect of temperature, ionic strength and surfactant on cellulase adsorption on isolated lignin was also explored in this study. The thermodynamic analysis of enzyme adsorption onto isolated lignin (Gibbs free energy change ΔG ≈ -30 kJ/mol) indicated this adsorption was a spontaneous process. The addition of surfactant (0.2% w/v) could reduce the adsorption of cellulase onto CEL-SELP by 60%.  Two types of adsorption isotherm were compared for cellulase adsorption onto isolated lignin. Langmuir adsorption isotherm showed better fit for the experimental data than that from Freundlich adsorption isotherm