1887

Abstract

Summary

Virus-specific antigens in cells infected with herpes simplex virus were separated on a preparative scale by two-stage polyacrylamide-gel electrophoresis. In the first stage, termed ‘electrofiltration’, the extract was electrophoresed through 3.5% gel to remove larger aggregates. Antigens in the resulting filtrate were then separated in the second stage by electrophoresis through 7% gel. The protein was recovered by sectioning the gel followed by overnight elution. Recoveries of about 70% were obtained from a 12 to 15 mg. load. Immunodiffusion and analytical electrophoresis both confirmed that separation had been achieved.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-4-2-151
1969-03-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/4/2/JV0040020151.html?itemId=/content/journal/jgv/10.1099/0022-1317-4-2-151&mimeType=html&fmt=ahah

References

  1. Andrews P. 1965; The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J 96:595
    [Google Scholar]
  2. Avrameas S., Ternynck T. 1967; Biologically active water-insoluble polymers. I. Their use for isolation of antigens and antibodies. J. biol. Chem 242:1651
    [Google Scholar]
  3. Bishop D. H. L., Claybrook J. R., Spiegelman S. 1967; Electrophoretic separation of viral nucleic acids on polyacrylamide gels. J. molec. Biol 26:373
    [Google Scholar]
  4. Brewer T. M. 1967; Artifact production in disc electrophoresis by ammonium persulphate. Science, N. Y 156:256
    [Google Scholar]
  5. Davis B. J. 1964; Disc electrophoresis. II. Method and application to serum proteins. Ann. N. Y. Acad. Sci 121:404
    [Google Scholar]
  6. Gibbs A. J., Harrison B. D., Watson D. H., Wildy P. 1966; What’s in a virus name ?. Nature, Lond 209:450
    [Google Scholar]
  7. Ginsberg H. S., Pereira H. G., Valentine R. C., Wilcox W. C. 1966; A proposed terminology for the adenovirus antigens and virion morphological subunits. Virology 28:782
    [Google Scholar]
  8. Heideman M. L. 1964; Disc electrophoresis of 131I-Iabelled protein hormone preparations and their reaction product with antibodies. Ann. N,. Y. Acad. Sci 121:501
    [Google Scholar]
  9. Jovin T., Chrambach A., Naughton M. A. 1964; An apparatus for preparative temperature-regulated polyacrylamide gel electrophoresis. Amlyt. Biochem 9:351
    [Google Scholar]
  10. Keir H. M. 1968; Virus induced enzymes in mammalian cells. In The Molecular Biology of Viruses. Symp. Soc. gen. Microbiol. no 18 Ed Crawford L. V., Stoker M. G. P. Cambridge University Press:
    [Google Scholar]
  11. Love R., Wildy P. 1963; Cytochemical studies on the nucleoproteins of HeLa cells infected with herpes virus. J. Cell Biol 17:237
    [Google Scholar]
  12. Lowry D. H., Roseborough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin-phenol reagent. J. biol. Chem 193:265
    [Google Scholar]
  13. Mitchell W. M. 1967; A potential source of electrophoretic artifacts in polyacrylamide gels. Biochim. biophys. Acta 147:171
    [Google Scholar]
  14. Ornstein L. 1964; Disc electrophoresis. I. Background and theory. Ann. N. Y. Acad. Sci 131:321
    [Google Scholar]
  15. Spiegel M., Ozaki M., Tyler A. 1965; Electrophoretic examination of soluble proteins synthesised in early sea urchin development. Biochem. biophys. Res. Commun 21:135
    [Google Scholar]
  16. Stephen J., Gallop R. G. C., Smith H. 1966; Separation of antigens by immunogenicity. Use of disulphide linked antibodies as immunosorbents. Biochem. J 101:717
    [Google Scholar]
  17. Sulitzeanu D., Slavin M., Yecheskeli E. 1967; Simplified technique for preparative disc electrophoresis. II. Further improvements in apparatus and some details of performance. Analyt. Biochem 21:57
    [Google Scholar]
  18. Watson D. H., Wildy P. 1969; The preparation of ‘monoprecipitin’ antisera to herpes virus specific antigens. J. gen. Virol 4:163
    [Google Scholar]
  19. Watson D. H., Wildy P., Russell W. C. 1964; Quantitative electron microscopic studies on the growth of herpes virus using the techniques of negative staining and ultramicrotomy. Virology 19:250
    [Google Scholar]
  20. Watson D. H., Shedden W. I. H., Elliot A., Tetsuka T., Wildy P., Bourgaux-Ramoisy D., Gold E. 1966; Virus specific antigens in mammalian cells infected with herpes simplex virus. Immunology 11:399
    [Google Scholar]
  21. Wright G. L., Mallmann W. L. 1966; Differential disc electrophoresis of serum proteins. Proc. Soc. exp. Biol. Med 123:22
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-4-2-151
Loading
/content/journal/jgv/10.1099/0022-1317-4-2-151
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error