Two small antigens have been isolated from suspensions of vesicular stomatitis virus grown in baby hamster kidney cells. The suspensions were first centrifuged at 15,000 × G for 100 min to remove the virion and other large antigens and the supernatant was then centrifuged at 80,000 × G for 180 min. This second supernatant was further fractionated by prolonged centrifugation in a sucrose density gradient and by filtration through Sephadex G-200. The antigens separated by these procedures stimulate the production of neutralizing antibodies in guinea pigs.

The approximate size of the antigens has been estimated by measuring their rates of sedimentation in sucrose gradients and observing their elution pattern in Sephadex G-200. The larger of the two antigens has an S value intermediate between those of 7S γ-globulin and bovine plasma albumin. The smaller antigen sediments more slowly than albumin. Similarly, the larger of the two antigens was eluted from Sephadex G-200 after 7S γ-globulin but before albumin and the smaller antigen was eluted after the albumin.

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