Identification of Immunoreactive Viral Proteins

  • Walter Hinderer
  • Bodo Plachter
  • Rolf Vornhagen
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 33)


Several diagnostic tools are available for the identification of acute and latent viral infections. Although newly developed nucleic acid amplification methods, such as the polymerase chain reaction (PCR), have proved to be very useful diagnostic procedures, conventional methods, such as cell culture and serology, still play an important role in viral diagnostics. Despite the fact that modern serological assays, such as enzyme-linked immunosorbent assay (ELISA), are inexpensive and easy to perform, there is a strong demand to improve the performance of such systems. Most serological tests are based on poorly characterized antigens produced in infected culture cells. It has been shown, however, that only few viral antigens contained in these preparations are essential for serodiagnosis. In addition, numerous viral proteins display homologies with their counterparts from related viruses. Finally, the specificity of serological assays can also be reduced by contaminating proteins from host cells. Selective purification of natural viral antigens using, for example, immunoaffinity chromatography is one possible way to improve the quality of an antibody assay. However, the low concentration of most viral proteins in cell culture-derived antigen preparations reduces the practicability of this approach.


Sodium Dodecyl Sulfate Viral Antigen Polymerase Chain Reaction Primer Solubilization Buffer Refold Buffer 
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  1. 1.
    Britt, W. J. and Alford, C. A. (1996) Cytomegalovirus, in Virology (Fields, B. N., Knipe, D. M., and Howley, P. M., eds.), Lippincott-Raven, Philadelphia, pp. 2493–2524.Google Scholar
  2. 2.
    Landini, M. P. and Mach, M. (1995) Searching for antibodies specific for human cytomegalovirus: is it diagnostically useful? When and how. Scand. J. Infect. Dis. Suppl. 99, 18–23.PubMedGoogle Scholar
  3. 3.
    Smith, D. B. and Johnson, K. S (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusion with glutathione S-transferase. Gene 67, 31–40.CrossRefPubMedGoogle Scholar
  4. 4.
    Frangioni, J. V. and Neel, B. G. (1993) Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. Anal. Biochem. 210, 179–187.CrossRefPubMedGoogle Scholar
  5. 5.
    Marston, F. A. (1986) The purification of eukaryotic polypeptides synthesized in Escherichia coli. Biochem. J. 240, 1–12.PubMedGoogle Scholar
  6. 6.
    Ghosh, S., Basu, S., Strum, J. C., and Bell, R. M. (1995) Identification of conditions that facilitate the expression of GST fusions as soluble, full-length proteins. Anal. Biochem. 225, 376–378.CrossRefPubMedGoogle Scholar
  7. 7.
    Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.CrossRefPubMedGoogle Scholar
  8. 8.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning—A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar

Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Walter Hinderer
    • 1
  • Bodo Plachter
    • 2
  • Rolf Vornhagen
    • 1
  1. 1.Research DepartmentBiotest AGDreieichGemany
  2. 2.Institute for VirologyUniversity of MainzMainzGermany

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