Isolation of Bacterial H-Antigens

Abstract

The cells of certain bacterial species are motile; they are able to penetrate semisolid agar layers. This motility of cells frequently occurs in Gram-negative species and is due to the existence of flagella on the bacterial cells. These hair-like flagella are highly antigenic and, since the thorough studies of Craigie (1931), it is generally accepted that H-antigenic properties of certain bacteria are associated with the flagella. The nature of flagellar antigens was reviewed by Koffler and Smith (1971). Weibull (1949a, 1949b) showed that the flagella consist of fibrous proteins and can be purified by fractional sedimentation in a high-speed centrifuge and also by ammonium sulfate precipitation. In the method described here, motile bacteria will be selected according to the procedure of Craigie. The flagella will be separated from the cells and their fibrous protein content, called flagellin, will be partially purified. The method described here is a modification of the descriptions of Weibull and also of Koffler and Kobayashi (1957).