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. 1992 Nov;30(11):2874–2880. doi: 10.1128/jcm.30.11.2874-2880.1992

Baculovirus expression of the nucleoprotein gene of measles virus and utility of the recombinant protein in diagnostic enzyme immunoassays.

K B Hummel 1, D D Erdman 1, J Heath 1, W J Bellini 1
PMCID: PMC270545  PMID: 1452657

Abstract

A recombinant baculovirus that expresses the nucleoprotein gene of measles virus (Edmonston vaccine strain) under the transcriptional control of the polyhedrin promoter was generated. The expressed protein (B-MVN) comigrated with the authentic viral nucleoprotein as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was phosphorylated. The B-MVN protein proved to be reactive with monoclonal antibodies in radioimmunoprecipitations, and it was immunogenic, eliciting in mice antisera that recognized the native nucleoprotein. In addition, the B-MVN protein was evaluated as a replacement source of antigen for whole virus in enzyme immunoassays (EIAs) for detection of measles virus-specific immunoglobulin M (IgM) and IgG antibodies. A capture IgM EIA with the B-MVN protein as antigen detected specific IgM antibodies in 18 (72%) acute- and all convalescent-phase specimens from 25 clinical measles cases and exceeded 99% specificity with 120 control specimens. An indirect IgG EIA with the B-MVN protein detected specific IgG antibodies in 129 of 131 (98%) serum specimens with antibodies to measles virus, and results obtained from testing 268 additional serum specimens were better correlated with measles virus-neutralizing antibodies than those obtained with a commercial EIA.

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Selected References

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

  1. Albrecht P., Herrmann K., Burns G. R. Role of virus strain in conventional and enhanced measles plaque neutralization test. J Virol Methods. 1981 Dec;3(5):251–260. doi: 10.1016/0166-0934(81)90062-8. [DOI] [PubMed] [Google Scholar]
  2. Anderson L. J., Hierholzer J. C., Stone Y. O., Tsou C., Fernie B. F. Identification of epitopes on respiratory syncytial virus proteins by competitive binding immunoassay. J Clin Microbiol. 1986 Mar;23(3):475–480. doi: 10.1128/jcm.23.3.475-480.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker S. C., Moyer S. A. Encapsidation of Sendai virus genome RNAs by purified NP protein during in vitro replication. J Virol. 1988 Mar;62(3):834–838. doi: 10.1128/jvi.62.3.834-838.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bankamp B., Brinckmann U. G., Reich A., Niewiesk S., ter Meulen V., Liebert U. G. Measles virus nucleocapsid protein protects rats from encephalitis. J Virol. 1991 Apr;65(4):1695–1700. doi: 10.1128/jvi.65.4.1695-1700.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bellini W., Englund G., Rammohan K., McFarlin D. E. Evaluation of the structural proteins of the hamster neurotropic strain of measles virus with monoclonal antibodies. J Neuroimmunol. 1986 Apr;11(2):149–163. doi: 10.1016/0165-5728(86)90116-5. [DOI] [PubMed] [Google Scholar]
  6. Chen R. T., Markowitz L. E., Albrecht P., Stewart J. A., Mofenson L. M., Preblud S. R., Orenstein W. A. Measles antibody: reevaluation of protective titers. J Infect Dis. 1990 Nov;162(5):1036–1042. doi: 10.1093/infdis/162.5.1036. [DOI] [PubMed] [Google Scholar]
  7. Erdman D. D., Anderson L. J., Adams D. R., Stewart J. A., Markowitz L. E., Bellini W. J. Evaluation of monoclonal antibody-based capture enzyme immunoassays for detection of specific antibodies to measles virus. J Clin Microbiol. 1991 Jul;29(7):1466–1471. doi: 10.1128/jcm.29.7.1466-1471.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giraudon P., Jacquier M. F., Wild T. F. Antigenic analysis of African measles virus field isolates: identification and localisation of one conserved and two variable epitope sites on the NP protein. Virus Res. 1988 May;10(2-3):137–152. doi: 10.1016/0168-1702(88)90011-1. [DOI] [PubMed] [Google Scholar]
  9. Graves M., Griffin D. E., Johnson R. T., Hirsch R. L., de Soriano I. L., Roedenbeck S., Vaisberg A. Development of antibody to measles virus polypeptides during complicated and uncomplicated measles virus infections. J Virol. 1984 Feb;49(2):409–412. doi: 10.1128/jvi.49.2.409-412.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. J Gen Virol. 1987 May;68(Pt 5):1233–1250. doi: 10.1099/0022-1317-68-5-1233. [DOI] [PubMed] [Google Scholar]
  12. Norrby E., Orvell C., Vandvik B., Cherry J. D. Antibodies against measles virus polypeptides in different disease conditions. Infect Immun. 1981 Dec;34(3):718–724. doi: 10.1128/iai.34.3.718-724.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Orenstein W. A., Albrecht P., Herrmann K. L., Bernier R., Bart K. J., Rovira E. Z. The plaque-neutralization test as a measure of prior exposure to measles virus. J Infect Dis. 1987 Jan;155(1):146–149. doi: 10.1093/infdis/155.1.146. [DOI] [PubMed] [Google Scholar]
  14. Rozenblatt S., Eizenberg O., Ben-Levy R., Lavie V., Bellini W. J. Sequence homology within the morbilliviruses. J Virol. 1985 Feb;53(2):684–690. doi: 10.1128/jvi.53.2.684-690.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. WELLER T. H., COONS A. H. Fluorescent antibody studies with agents of varicella and herpes zoster propagated in vitro. Proc Soc Exp Biol Med. 1954 Aug-Sep;86(4):789–794. doi: 10.3181/00379727-86-21235. [DOI] [PubMed] [Google Scholar]

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