CHEMICAL STUDIES ON CELLULAR COMPONENTS OFBORDETELLA PERTUSSISI

Abstract

, AND YUICHI YAMAMURA. Chemical studies on cellular components of Bordete77a pertussis. I. Purification and properties of agglutinogen. J. Bacteriol. 82:648-656. 1961.-A method is described for the preparation of the agglutinogen of Hemophilus pertussis in phase I by methanol precipitation and ion-exchange chromatography. High homogeneity of this agglutinogen was evidenced by chromatography, but a trace of impurity was detected by ultracentrifugal analysis and by an agar gel diffusion test.

Purified K‐Agglutinogen of Bordetella pertussis and Its Properties

“…By gel-diffusion tests with anti-L and anti-S factor sera, we confirmed that the K-agglutinogen fraction CM-F4 consisted of both L and S antigens, and that CM-F5 consisted of mainly S antigen. Onoue et al [15], Schuchardt et al [19] and the present authors [11,12] recognized the K-agglutinogen as a simple protein. In the present paper, the fact was reconfirmed by using further purified K-agglutinogen.…”

“…Agglutinogen purified by Schuchardt et al [19] was slightly contaminated with protective antigen. Ishii [6] described the presence of protective activity in the agglutinogen obtained by Onoue et al [15]. We found that acid-heating, followed by salt or alcohol precipitation procedures before applying chromatography, made it difficult to separate clearly the denatured heat-labile toxin and other cell-components from the agglutinogen.…”

“…Smolens and Mudd [17], Zen-yoji [21], and Onoue et al [15] extracted K-agglutinogen by acid-heating. It is considered that the heat-labile agglutinogens such as the L-antigen might be denatured or destroyed during acid-heat extraction.…”

“…Attempts have been made to purify agglutinogen from Bordetella pertussis organisms by several workers [4,5,15,17,19,21]. Flosdorf and Kimball [4] separated the agglutinogen by isoelectric and ammonium sulfate precipitations from the supernatant of ultrasonic-treated cells.…”

“…Both agglutinogens obtained by Smolens and Mudd, and Zen-yoji were non-necrotic and efficient as skin test antigens. Onoue et al [15] at-tempted to purify the agglutinogen by a modified Smolens and Mudd's method followed by column chromatography on Odiethylaminoethyl (DEAE)-cellulose, and Schuchardt et al [19] separated the agglutinogen by sodium sulfate fractionation from the supernatant of sonic treated cells. These workers demonstrated the agglutinogen to be a simple protein free from heatlabile toxin.…”