Radioimmunoassay for the antigenic determinants of cholera toxin and its components

K. E. Hejtmancik, Johnny Peterson, D. E. Markel, A. Kurosky

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Abstract

A radioimmunoassay procedure is described for the detection of cholera toxin and its component polypeptide chains. Cholera toxin, A subunit, B subunit, α chain, and γ chain were iodinated by the chloramine T procedure. Radiolabeling did not significantly alter the polyacrylamide electrophoretic migration patterns of the toxin or its components. Moreover, radiolabeled toxin, B subunit, and α chain preparations retained substantial ability to bind to intestinal mucosal homogenates. The minimal amount of antitoxin detectable with radiolabeled toxin was 0.04 antitoxin units/ml. Substitution of radiolabeled B subunit, A subunit, and α chain for radiolabeled toxin decreased the sensitivity of the test. Radiolabeled γ chain did not bind to the antitoxin preparation. Competitive inhibition studies, with titrated anti-choleragen serum and radiolabeled toxin or components, indicated that the minimum concentration of toxin detectable was 7.0 x 10-8 μmol/ml at a 90% inhibition level. The A subunit and α chain preparations inhibited the binding of the radiolabeled B subunit to antitoxin sites. Conversely, B subunit inhibited the binding of radiolabeled A subunit and α chain to antitoxin. The γ chain did not show any reaction with antitoxin or cross-reaction with either whole toxin or its components. These results strongly suggest that the A subunit and the α chain contain antigenic determinant(s) that are common to the B subunit. The B subunit (β chain) and the α chain of cholera toxin may therefore contain region(s) of chemical similarity.

Original languageEnglish (US)
Pages (from-to)621-628
Number of pages8
JournalInfection and Immunity
Volume17
Issue number3
StatePublished - 1977

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Antitoxins
Cholera Toxin
Radioimmunoassay
Epitopes
Cross Reactions
Peptides
Serum

ASJC Scopus subject areas

  • Immunology

Cite this

Radioimmunoassay for the antigenic determinants of cholera toxin and its components. / Hejtmancik, K. E.; Peterson, Johnny; Markel, D. E.; Kurosky, A.

In: Infection and Immunity, Vol. 17, No. 3, 1977, p. 621-628.

Research output: Contribution to journalArticle

Hejtmancik, KE, Peterson, J, Markel, DE & Kurosky, A 1977, 'Radioimmunoassay for the antigenic determinants of cholera toxin and its components', Infection and Immunity, vol. 17, no. 3, pp. 621-628.
Hejtmancik, K. E. ; Peterson, Johnny ; Markel, D. E. ; Kurosky, A. / Radioimmunoassay for the antigenic determinants of cholera toxin and its components. In: Infection and Immunity. 1977 ; Vol. 17, No. 3. pp. 621-628.
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