Abstract: An antigenic composition is derived from surface extracted protein of cell wall surfaces and the culture supernatant extract of Staphylococci. The strain is highly virulent and &bgr; hemolytic on blood agar plate. They are mixed and purified by ion exchange and gel filtration column chromatography. This preparation method can be used in all Gram-positive bacilli. The antigens, having molecular weight of about 10,000-70,000 are certain kinds of glycoprotein comprising proteins (ca. 10-20%) and carbohydrates (ca. 75-90%). They are extracted from the cells with the use of a hypertonic buffer solution of pH 6.5-8.5 at a temperature below the denaturing point of the antigenic protein, and are salted out with ammonium sulfate (ca. 65-85%). The culture supernatant is extracted in the same way by salting out with ammonium sulfate (ca. 65-85%). The antigen fraction from both cell wall and culture supernatant is obtained in such an operation. The antigen can be used as a preparation antigen to Staphylococcus antigen.