Publication: The study of intestinal immunity against V. Cholerae: Purification of V. cholerae El Tor haemagglutinin and the protective role of its antibody in experimental cholera
Issued Date
1979-12-01
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ISSN
00383619
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2-s2.0-0018634778
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Mahidol University
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SCOPUS
Bibliographic Citation
Southeast Asian Journal of Tropical Medicine and Public Health. Vol.10, No.1 (1979), 73-80
Suggested Citation
W. Chaicumpa, N. Atthasishtha The study of intestinal immunity against V. Cholerae: Purification of V. cholerae El Tor haemagglutinin and the protective role of its antibody in experimental cholera. Southeast Asian Journal of Tropical Medicine and Public Health. Vol.10, No.1 (1979), 73-80. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/13197
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Title
The study of intestinal immunity against V. Cholerae: Purification of V. cholerae El Tor haemagglutinin and the protective role of its antibody in experimental cholera
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Abstract
V. Cholerae El Tor Ogawa strain O 17 SR grown on trypticase soy agar were extracted with 0.05 M cyclohexylamino propane sulfonic acid (CAPS) pH 9.5 at 37°C for 1 hour. The bacteria were then removed by centrifugation and millipore filtration. The filtered fluid, after being dialysed against many changes of cold distilled water, was concentrated and passed through Sephadex G 200 column. Three protein profiles were eluted out with 0.05 M Tris buffer pH 8.6. The haemagglutinin and the bacterial lipopolysaccharide (LPS) were confined to the first profile. They were subsequently separated by agarose gel electrophoresis. The haemagglutinin was found to be more anodic than the LPS. After homogenization of the gel strips containing the haemagglutinin followed by centrifugation at 9,000 g pure haemagglutinin was obtained in the supernatant. Rabbit antiserum against pure haemagglutinin contained protective antibodies against V. cholerae infection in the baby mouse model. Specific antibodies prepared from this antiserum was as protective as the antibodies directed against whole V. cholerae and heat-stable somatic antigens of V. cholerae upon the same weight unit.