A series of novel magnetic microspheres were prepared from chitosan and β-cyclodextrin through inverse suspension crosslinking. The obtained magnetic microspheres were characterized by optics microscopy，IR and VSM. The adsorption behavior of papain on microspheres was studied also.The results showed that the microspheres were spherical shape and good magnetic response. the adsorption of papain on microspheres was described by both the Langmuir adsorption isotherm and the Freundlich isotherm. Sodium dodecyl sulfate-polyacrylamide gel exectrophoresis（SDS-PAGE）showed that papain was desorbed form magnetic microspheres without denaturation.