Studies were initiated in an attempt to produce an inactivated vaccine of high potency with low toxicity. In this study structural integrity of bacteria can be preserved when different radiation doses of gamma rays were used to inactivate S. aureus. The results revealed that the 7 KGy of γ- radiation was the most suitable dose for attenuation S. aureus. Moreover, this study demonstrated that exposure of S. aureus to γ- radiation at a dose of 7 KGy inactivated bacterial cells growth with low DNA damage and did not alter bacteria protein profile. Meanwhile, higher radiation doses caused an obvious DNA damage and alteration in the bacterial protein profile. Furthermore, the effect of gamma irradiated S. aureus on the innate and adaptive immune components after vaccination was demonstrated. All the measured immunological parameters (TLR2, IL10, IL1β, iNOs, TGFβ, MAPK and PI3K) were increased when compared to the control group. This approach could expedite vaccine production for emerging and established pathogens for which no protective vaccines exist.