1887

Abstract

Summary

Monoclonal antibody (MAb) to diphtheria toxin was produced in mouse hybridomas, and shown by ELISA to be of sub-class IgG. Hybridomas were inoculated into mice to produce ascitic fluid from which MAb was purified by caprylic acid. The MAb was shown by immunoblotting to be directed against the A fragment of the toxin and also against the intact toxin molecule. After conjugation with fluorescein isothiocyanate, it was used in an immunoassay to detect toxin in culture supernates of and . The assay correlated well with the Elek test and with virulence in guinea-pigs; but it gave occasional false positive results, probably by binding of MAb to defective toxin.

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1990-08-01
2024-05-02
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References

  1. Davies JR. Elek’s test for the toxigenicity of Corynebacteria. In: Davies JR, Glencross EJG, Marks J, Plow CD, Thomas MEM. Laboratory Methods 1. Public Health Laboratory Service Monograph Series no. 5 London: HMSO; 19741–7
    [Google Scholar]
  2. Snell JJS, DeMello JV, Gardner PS, Kwantes W, Brooks R. Detection of toxin production by Corynebacterium diphtheriae : results of a trial organised as part of the United Kingdom National External Microbiological Quality Assessment Scheme. J Clin Pathol 1984; 37:796–799
    [Google Scholar]
  3. Russell WC, Patel G, Precious B, Sharp I, Gardner PS. Monoclonal antibodies against Adenovirus Type 5: preparation and preliminary characterization. J Gen Virol 1981; 56:393–408
    [Google Scholar]
  4. Tedder RS, Yao JL, Anderson MJ. The production of monoclonal antibodies to rubella haemagglutinin and their use in antibody-capture assays for rubella-specific IgM. JHyg (Lond) 1982; 88:335–350
    [Google Scholar]
  5. Steinbuch M, Audran R. The isolation of IgG from mammalian sera with the aid of caprylic acid. Arch Biochem Biophys 1969; 134:279–284
    [Google Scholar]
  6. Samuel D, Patt RJ, Abuknesha RA. A sensitive method of detecting proteins on dot and Western blots using a monoclonal antibody to FITC. J Immunol Methods 1988; 107:217–224
    [Google Scholar]
  7. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
    [Google Scholar]
  8. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
    [Google Scholar]
  9. Leary JJ, Brigati DJ, Ward DC. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci USA 1983; 80:4045–4049
    [Google Scholar]
  10. Hallas G. The use of SDS-polyacrylamide gel electrophoresis in epidemiological studies of Corynebacterium diphtheriae. Epidemiol Infect 1988; 100:83–90
    [Google Scholar]
  11. Krech T, Wittelsburger C. Immunologische Methoden zum Nachweis von Diphtherie-Toxin (Passive Haemagglutionation und ELISA zum Toxinnachweis aus Kulturen und im Serum). Zentralbl Bakteriol Mikrobiol Hyg (A) 1987; 265:124–135
    [Google Scholar]
  12. Sottnek FO, Miller JM. Isolation and identification of Corynebacterium diphtheriae. US Department of Health and Human Services. Atlanta, Public Health Service, Center for Disease Control 19801–14
    [Google Scholar]
  13. Murphy JR. The diphtheria toxin structural gene. Curr Top Microbiol Immunol 1985; 118:235–251
    [Google Scholar]
  14. Communicable disease statistics Office of Population Censuses and Surveys, OPCS series, MB2 no. 13. London: HMSO; 19866
    [Google Scholar]
  15. Bjorkholm B, Oiling S, Larsson P, Hagberg L. An outbreak of diphtheria among Swedish alcoholics. Infection 1987; 15:354–358
    [Google Scholar]
  16. Karzon DT, Edwards KM. Diphtheria outbreaks in immunized populations (Editorial). NEngl J Med 1988; 318:41–43
    [Google Scholar]
  17. Sheffield FW. (Ad-hoc Working Group). Susceptibility to diphtheria. Lancet 1978; 1:428–430
    [Google Scholar]
  18. Warwick-Brown NP, Lund VJ. Diphtheria—a forgotten disease. Br J Clin Prac 1987; 41:580–582
    [Google Scholar]
  19. Simmons LE, Abbott JD, Macaulay ME et al. Diphtheria carriers in Manchester: simultaneous infection with toxigenic and non-toxigenic mitis strains. Lancet 1980; 1:304–305
    [Google Scholar]
  20. Pappenheimer AM, Johnson SJ. Studies in diphtheria toxin production. I: the effect of iron and copper. Br J Exp Pathol 1936; 17:335–341
    [Google Scholar]
  21. Ionescu M, Dumitrescu A, Stefanescu M, Szegli G. ELISA for toxinogenesis identification in Corynebacterium diphtheriae, diagnostic value. Arch Roum Pathol Exp Microbiol 1986; 45:5–12
    [Google Scholar]
  22. Nielsen PB, Koch C, Friis H, Heron I, Prag J, Schmidt J. Double-antibody sandwich enzyme-linked immunosorbent assay for rapid detection of toxin-producing Corynebacterium diphtheriae. J Clin Microbiol 1987; 25:1280–1284
    [Google Scholar]
  23. Barksdale L. Corynebacterium diphtheriae and its relatives. Bacteriol Rev 1970; 34:378–422
    [Google Scholar]
  24. Hayakawa S, Uchida T, Mekada E, Moynihan MR, Okada Y. Monoclonal antibody against diphtheria toxin— effect on toxin binding and entry into cells. J Biol Chem 1983; 258:4311–4317
    [Google Scholar]
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