Porous chitosan microspheres suitable for the delivery of antigen were prepared using a wet phase-inversion method. The pore structure of the chitosan microsphere could be modified by the change of pH value of the coagulation medium, which is the aqueous tripolyphosphate (TPP) solution. High porosity of chitosan microsphere with an open porous structure on its surface was prepared by coagulation in TPP aq. solution of pH 8.9. The porous chitosan microspheres were modified chemically with reagents to introduce three types of functional groups; carboxyl, hydrophobic acyl and quaternary ammonium groups. Antigen of ND vaccine was immobilized into the pores of porous chitosan microspheres and the adsorbed antigen was assayed by the Hemoglobin Aggregation (HA) analytical method. Sustained-release of ND vaccine's antigen could be achieved through an adsorption-desorption release test. The chemical modifications of the porous chitosan microspheres have a strong large influence on the adsorption efficiency or release rates of the antigen investigated. The porous microspheres have a higher adsorption efficiency and the slower release rate of antigen when modified chemically with 3-chloro-2-hydroypropyltrimethylamonium chloride.