The aim of this study was to prepare poly(lactic-co-glycolic acid) (PLGA) microspheres containing hepatitis B surface antigen (HBsAg) using different stabilizer such as Sucrose, Trehalose, Bovine serum albumin (BSA), Human serum albumin (HSA), Hydroxypropyl-b-Cyclodextrin (HP-b-CD) and combination of Trehalose and HP-b-CD. Suitable stabilizer is required in the PLGA based formulation containing HBsAg to protect the HBsAg antigenicity loss due to usage of organic solvents, release of acids from PLGA and lyophilization. The developed formulations were characterized for entrapment efficiency, antigen structural integrity, Invitro release study, surface morphology and particle size. The entrapment efficiency observed more with trehalose, HP-b-CD and its combination. There is no major structural antigen integrity by SDS-PAGE for formulations with different stabilizers. The in vitro release test showed that combination of Trehalose and HP-b-CD in both primary and secondary emulsion shows improved antigen recovery to >90% on 42 days release as compared to other formulation. Based on these findings, the HBsA with trehalose and HP-b-CD loaded PLGA microspheres could be an effective carrier delivery system for HBsAg