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. 1994 Jan;1(1):78–82. doi: 10.1128/cdli.1.1.78-82.1994

Enzyme-linked immunosorbent assay using a recombinant baculovirus-expressed Bacillus anthracis protective antigen (PA): measurement of human anti-PA antibodies.

L C Iacono-Connors 1, J Novak 1, C Rossi 1, J Mangiafico 1, T Ksiazek 1
PMCID: PMC368200  PMID: 7496927

Abstract

We developed an antigen capture enzyme-linked immunosorbent assay (ELISA) which does not require purified protective antigen (PA) for detection of human antibodies to Bacillus anthracis PA. Lysates of Spodoptera frugiperda (Sf-9) cells infected with recombinant baculovirus containing the PA gene were used as the source of PA to develop the ELISA. Recombinant PA from crude Sf-9 cell lysates or PA purified from B. anthracis Sterne strain was captured by an anti-PA monoclonal antibody coated onto microtiter plates. We demonstrated that human serum antibody titers to PA were identical in the ELISA whether we used crude Sf-9 cell lysates containing recombinant baculovirus-expressed PA or purified Sterne PA. Finally, false-positive results observed in a direct ELISA were eliminated with this antigen capture ELISA. Thus, the antigen capture ELISA with crude preparations of baculovirus-expressed PA is reliable, safe, and inexpensive for determining anti-PA antibody levels in human sera.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brachman P. S., Gold H., Plotkin S. A., Fekety F. R., Werrin M., Ingraham N. R. Field Evaluation of a Human Anthrax Vaccine. Am J Public Health Nations Health. 1962 Apr;52(4):632–645. doi: 10.2105/ajph.52.4.632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Buchanan T. M., Feeley J. C., Hayes P. S., Brachman P. S. Anthrax indirect microhemagglutination test. J Immunol. 1971 Dec;107(6):1631–1636. [PubMed] [Google Scholar]
  3. Fish D. C., Mahlandt B. G., Dobbs J. P., Lincoln R. E. Purification and properties of in vitro-produced anthrax toxin components. J Bacteriol. 1968 Mar;95(3):907–918. doi: 10.1128/jb.95.3.907-918.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hambleton P., Carman J. A., Melling J. Anthrax: the disease in relation to vaccines. Vaccine. 1984 Jun;2(2):125–132. doi: 10.1016/0264-410x(84)90003-3. [DOI] [PubMed] [Google Scholar]
  5. Iacono-Connors L. C., Schmaljohn C. S., Dalrymple J. M. Expression of the Bacillus anthracis protective antigen gene by baculovirus and vaccinia virus recombinants. Infect Immun. 1990 Feb;58(2):366–372. doi: 10.1128/iai.58.2.366-372.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ivins B. E., Welkos S. L., Little S. F., Crumrine M. H., Nelson G. O. Immunization against anthrax with Bacillus anthracis protective antigen combined with adjuvants. Infect Immun. 1992 Feb;60(2):662–668. doi: 10.1128/iai.60.2.662-668.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ivins B. E., Welkos S. L. Recent advances in the development of an improved, human anthrax vaccine. Eur J Epidemiol. 1988 Mar;4(1):12–19. doi: 10.1007/BF00152686. [DOI] [PubMed] [Google Scholar]
  8. Johnson-Winegar A. Comparison of enzyme-linked immunosorbent and indirect hemagglutination assays for determining anthrax antibodies. J Clin Microbiol. 1984 Sep;20(3):357–361. doi: 10.1128/jcm.20.3.357-361.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Leppla S. H. Production and purification of anthrax toxin. Methods Enzymol. 1988;165:103–116. doi: 10.1016/s0076-6879(88)65019-1. [DOI] [PubMed] [Google Scholar]
  10. Little S. F., Knudson G. B. Comparative efficacy of Bacillus anthracis live spore vaccine and protective antigen vaccine against anthrax in the guinea pig. Infect Immun. 1986 May;52(2):509–512. doi: 10.1128/iai.52.2.509-512.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Little S. F., Leppla S. H., Cora E. Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin. Infect Immun. 1988 Jul;56(7):1807–1813. doi: 10.1128/iai.56.7.1807-1813.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Little S. F., Leppla S. H., Friedlander A. M. Production and characterization of monoclonal antibodies against the lethal factor component of Bacillus anthracis lethal toxin. Infect Immun. 1990 Jun;58(6):1606–1613. doi: 10.21236/ada216203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meegan J. M., Yedloutschnig R. J., Peleg B. A., Shy J., Peters C. J., Walker J. S., Shope R. E. Enzyme-linked immunosorbent assay for detection of antibodies to Rift Valley fever virus in ovine and bovine sera. Am J Vet Res. 1987 Jul;48(7):1138–1141. [PubMed] [Google Scholar]
  14. PUZISS M., MANNING L. C., LYNCH J. W., BARCLAYE, ABELOW I., WRIGHT G. G. Large-scale production of protective antigen of Bacillus anthracis in anaerobic cultures. Appl Microbiol. 1963 Jul;11:330–334. doi: 10.1128/am.11.4.330-334.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Turnbull P. C., Broster M. G., Carman J. A., Manchee R. J., Melling J. Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity. Infect Immun. 1986 May;52(2):356–363. doi: 10.1128/iai.52.2.356-363.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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