The adsorption equilibrium and spacer arm effect based on the adsorption of asparaginase on activated sepharose 4B with aliphatic diamines (1,4-diaminebutane-1,8-diamineoctane) as the spacer arms and L(+)chlorosuccinamic acid as the ligand was studied in a batch reactor and in packed bed. The adsorption equilibrium constants were determined for all the adsorbents. A maximum adsorption was observed when 1,6 diaminehexane was used as spacer arm, under optimum adsorption conditions. iPartition coefficient was also determined. This coefficient allows us the calculation of the adsorption equilibrium constant inside the particles. A pulse-response chromatographic method and statistical moments were used to determine the adsorption equilibrium constants in the packed bed. The values of the adsorption equilibrium constants for different adsorbents were compared with those obtained previously in the batch reactor taking into account the mass-transfer resistance inside the particles of the adsorbent (partition coefficient). The difference in the values obtained was lower than 5-8%.