1887

Abstract

SUMMARY

When the polypeptides of hepatitis B surface antigen were examined by SDS-polyacrylamide gel electrophoresis under a variety of conditions, anomalous results were found to be due to (i) variable and at times incomplete dissociation of polypeptides after boiling with 1% SDS and reducing agent, (ii) reaggregation of solubilized material under certain electrophoretic conditions and during laboratory manipulations, and (iii) the variable presence of additional components in hepatitis B surface antigen prepared from certain individual donors. When these factors were taken into account, two major components were consistently identified by discontinuous buffer polyacrylamide gel electrophoresis, of apparent mol. wt. 60000 to 70000 and 12000 to 14000. However, in view of the demonstrated limitations of this technique in examining HBAg polypeptides, alternative methods are necessary to confirm the true mol. wt. of the unique virus-specified amino acid sequence present.

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1976-11-01
2024-04-24
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References

  1. Burrell C. J. 1975; Host components in hepatitis B antigen. Journal of General Virology 27:117–126
    [Google Scholar]
  2. Burrell C. J., Leadbetter O., Mackay P., Marmion B. P. 1976; Tryptic cleavage of antibody binding sites from hepatitis B surface antigen. Journal of General Virology 33:41–50
    [Google Scholar]
  3. Camacho A., Carraseosa J. L., Vinuela E., Salas M. 1975; Discrepancy in the mobility of a protein of phage ɸ 29 in two different SDS polyacrylamide-gel systems. Analytical Biochemistry 69:395–400
    [Google Scholar]
  4. Chairez R., Steiner S., Melnick J. L., Dreesman G. R. 1973; Glycoproteins associated with hepatitis B antigen. Intervirology 1:224–228
    [Google Scholar]
  5. Chairez R., Hollinger F. B., Brunschwig J. P., Dreesman G. R. 1975; Comparative biophysical studies of hepatitis B antigen, subtypes adw and ayw. Journal of Virology 15:182–190
    [Google Scholar]
  6. Dane D. S., Cameron C. H., Briggs M. 1970; Virus-like particles in serum of patients with Australia antigen-associated hepatitis. Lancet i:695–698
    [Google Scholar]
  7. Dreesman G. R., Hollinger F. B., Suriano J. R., FujiokA R. S., Brunschwig J. P., Melnick J. L. 1972; Biophysical and biochemical heterogeneity of purified hepatitis B antigen. Journal of Virology 10:469–476
    [Google Scholar]
  8. Dreesman G. R., Chairez R., Suarez M., Hollinger F. B., Courtney R. J., Melnick J. L. 1975; Production of antibody to individual polypeptides derived from purified hepatitis B surface antigen. Journal of Virology 16:508–515
    [Google Scholar]
  9. Fairbanks G., Steck T. L., Wallach D. F. H. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617
    [Google Scholar]
  10. Frank R. N., Rodbard D. 1975; Precision of sodium dodecyl sulphate-polyacrylamide-gel electrophoresis for the molecular weight estimation of a membrane glycoprotein: studies on bovine rhodopsin. Archives of Biochemistry and Biophysics 171:1–13
    [Google Scholar]
  11. Gerin J. L., Holland P. V., Purcell R. H. 1971; Australia antigen: large-scale purification from human serum and biochemical studies on its proteins. Journal of Virology 7:569–576
    [Google Scholar]
  12. Gerin J. L. 1972 In Hepatitis and Blood Transfusion pp 205–219 Edited by Vyas G. N., Perkins H. A., Schmid R. S. New York and London: Grune & Stratton;
    [Google Scholar]
  13. Gerlich W., May G. 1973; Hepatitis assoziiertes Antigen: Reinigung und Eigenschaften. Zentralblatt für Bakteriologie, I. Abteilung Originate 224:49–60
    [Google Scholar]
  14. Green J. R., Dunn M. J., Spooner R. L., Maddy A. H. 1974; The immunological heterogeneity of the proteins extracted from Bos laurus erythrocyte membranes by EDTA. Biochimica et Biophysica Acta 373:51–57
    [Google Scholar]
  15. Howard C. R., Zuckerman A. J. 1974; Characterisation of hepatitis B antigen polypeptides. Intervirology 4:129–156
    [Google Scholar]
  16. Katzman R. L. 1971; The inadequacy of sodium dodecyl sulphate as a dissociative agent for brain proteins and glycoproteins. Biochimica et Biophysica Acta 226:269–72
    [Google Scholar]
  17. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  18. Maddy A. H., Dunn M. J. 1973 In Protides of the Biological Fluids vol 21 pp 21–26 Edited by Peeters H. Oxford, New York, Toronto, Sydney: Pergamon Press;
    [Google Scholar]
  19. Maizel J. V. 1969 In Fundamental Techniques in Virology pp 334–362 Edited by Hable K., Salzman N. P. New York and London: Academic Press;
    [Google Scholar]
  20. Morell P., Wiggins R. C., Gray M. J. 1975; Polyacrylamide gel electrophoresis of myelin proteins: a caution. Analytical Biochemistry 68:148–154
    [Google Scholar]
  21. Ornstein L. 1964; Disc electrophoresis-I. Background and theory. Annals of New York Academy of Science 121:321–349
    [Google Scholar]
  22. Overby L. R., Hung P. P., Mao J. C.-H., Ling C. M. 1975; Rolling circular DNA associated with Dane particles in hepatitis B virus. Nature, London 255:84–85
    [Google Scholar]
  23. Robinson W. S., Clayton D. A., Greenman R. L. 1974; DNA of a human hepatitis B candidate. Journal of Virology 14:384–391
    [Google Scholar]
  24. Shapiro A. L., Vinuela E., Maizel J. V. 1967; Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and Biophysical Research Communications 28:815–820
    [Google Scholar]
  25. Shih J. W.-K., Gerin J. L. 1975; Immunochemistry of hepatitis B surface antigen (HBsAg): preparation and characterization of antibodies to the constituent polypeptides. Journal of Immunology 115:634–639
    [Google Scholar]
  26. Swaney J. B., Van De Woude G. F., Bachrach H. L. 1974; Sodium dodecyl sulfate-dependent anomalies in gel electrophoresis: alterations in the banding patterns of foot-and-mouth disease virus polypeptides. Analytical Biochemistry 58:337–346
    [Google Scholar]
  27. Tanford C. 1968; Protein denaturation. Advances in Protein Chemistry 23:121–282
    [Google Scholar]
  28. Vyas G. N., Williams E. W., Klaus G. G. B., Bond H. E. 1972; Hepatitis-associated Australia antigen. Protein, peptides and amino acid composition of purified antigen with its use in determining sensitivity of the haemagglutination test. Journal of Immunology 108:1114–1118
    [Google Scholar]
  29. Weber K., Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulphate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
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