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. 1972 Sep;10(3):469–476. doi: 10.1128/jvi.10.3.469-476.1972

Biophysical and Biochemical Heterogeneity of Purified Hepatitis B Antigen

G R Dreesman 1,2, F B Hollinger 1,2, J R Suriano 1,2, R S Fujioka 1,2, J P Brunschwig 1,2, J L Melnick 1,2
PMCID: PMC356488  PMID: 4116138

Abstract

Hepatitis B antigen of the D (a+, d+, y−) subtype was purified from plasma of apparently healthy persons and from hepatitis patients. The original samples contained 20- and 42-nm particles and tubular forms (20-nm diameter). Ultracentrifugation during the purification procedure yielded pellets which were then treated at pH 2.4. Both the large, 42-nm Dane particles and the tubular forms were lost during the acid treatment of the pelleted particles, yielding a preparation containing a mixture of particles approximately 20 and 25 nm in diameter. This difference in size was substantiated in that two distinct molecular weights were calculated from high-speed equilibrium data, 3.6 × 106 and 4.5 × 106. Further heterogeneity was observed in that hepatitis B antigenic activity was present in purified particles with an isoelectric pH of 4.0 and also in those with a pH of 4.4. No significant differences were observed in the gross amino acid composition of purified antigen obtained from plasma of three different persons. 125I-labeled, purified antigen was found to contain six distinct polypeptides with molecular weights ranging from 10,000 to 39,000.

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Selected References

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  1. Almeida J. D., Rubenstein D., Stott E. J. New antigen-antibody system in Australia-antigen-positive hepatitis. Lancet. 1971 Dec 4;2(7736):1225–1227. doi: 10.1016/s0140-6736(71)90543-5. [DOI] [PubMed] [Google Scholar]
  2. Almeida J. D., Waterson A. P. The morphology of virus-antibody interaction. Adv Virus Res. 1969;15:307–338. doi: 10.1016/S0065-3527(08)60878-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BLUMBERG B. S. POLYMORPHISMS OF THE SERUM PROTEINS AND THE DEVELOPMENT OF ISO-PRECIPITINS IN TRANSFUSED PATIENTS. Bull N Y Acad Med. 1964 May;40:377–386. [PMC free article] [PubMed] [Google Scholar]
  4. Bayer M. E., Blumberg B. S., Werner B. Particles associated with Australia antigen in the sera of patients with leukaemia, Down's Syndrome and hepatitis. Nature. 1968 Jun 15;218(5146):1057–1059. doi: 10.1038/2181057a0. [DOI] [PubMed] [Google Scholar]
  5. Bond H. E., Hall W. T. Separation and purification of hepatitis-associated antigen into morphologic types by zonal ultracentrifugation. J Infect Dis. 1972 Mar;125(3):263–268. doi: 10.1093/infdis/125.3.263. [DOI] [PubMed] [Google Scholar]
  6. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  7. Dane D. S., Cameron C. H., Briggs M. Virus-like particles in serum of patients with Australia-antigen-associated hepatitis. Lancet. 1970 Apr 4;1(7649):695–698. doi: 10.1016/s0140-6736(70)90926-8. [DOI] [PubMed] [Google Scholar]
  8. Dreesman G. R., Hollinger F. B., McCombs R. M., Melnick J. L. Production of potent anti-Australia antigen sera of high specificity and sensitivity in goats. Infect Immun. 1972 Feb;5(2):213–221. doi: 10.1128/iai.5.2.213-221.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerin J. L., Holland P. V., Purcell R. H. Australia antigen: large-scale purification from human serum and biochemical studies of its proteins. J Virol. 1971 May;7(5):569–576. doi: 10.1128/jvi.7.5.569-576.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Greenberg H. B., Gocke D. J. An analysis of antibody response to Australia antigen in man. J Infect Dis. 1971 Apr;123(4):356–364. doi: 10.1093/infdis/123.4.356. [DOI] [PubMed] [Google Scholar]
  12. Gust I. D., Cross G., Kaldor J., Ferris A. A. Virus-like particles in Australia-antigen-associated hepatitis. Lancet. 1970 May 2;1(7653):953–953. doi: 10.1016/s0140-6736(70)91082-2. [DOI] [PubMed] [Google Scholar]
  13. Hollinger F. B., Vorndam V., Dreesman G. R. Assay of Australia antigen and antibody employing double-antibody and solid-phase radioimmunoassay techniques and comparison with the passive hemagglutination methods. J Immunol. 1971 Oct;107(4):1099–1111. [PubMed] [Google Scholar]
  14. Jenson A. B., McCombs R. M., Sakurada N., Melnick J. L. Organ cultures inoculated with serum from a hepatitis patient with Au antigenemia. Exp Mol Pathol. 1970 Oct;13(2):217–230. doi: 10.1016/0014-4800(70)90007-9. [DOI] [PubMed] [Google Scholar]
  15. Jokelainen P. T., Krohn K., Prince A. M., Finlayson N. D. Electrn microscopic observations on virus-like particles associated with SH antigen. J Virol. 1970 Nov;6(5):685–689. doi: 10.1128/jvi.6.5.685-689.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kim C. Y., Tilles J. G. Immunologic and electrophoretic heterogeneity of hepatitis-associated antigen. J Infect Dis. 1971 Jun;123(6):618–628. doi: 10.1093/infdis/123.6.618. [DOI] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Le Bouvier G. L., McCollum R. W. Australia (hepatitis-associated) antigen: physicochemical and immunological characteristics. Adv Virus Res. 1970;16:357–396. doi: 10.1016/S0065-3527(08)60027-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Le Bouvier G. L. The heterogeneity of Australia antigen. J Infect Dis. 1971 Jun;123(6):671–675. doi: 10.1093/infdis/123.6.671. [DOI] [PubMed] [Google Scholar]
  20. Levene C., Blumberg B. S. Additional specificities of Australia antigen and the possible identification of hepatitis carriers. Nature. 1969 Jan 11;221(5176):195–196. doi: 10.1038/221195a0. [DOI] [PubMed] [Google Scholar]
  21. McCombs R. M., Melnick M. B., Brunschwig J. P. Biophysical studies of vesicular stomatitis virus. J Bacteriol. 1966 Feb;91(2):803–812. doi: 10.1128/jb.91.2.803-812.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Raunio V. K., London W. T., Sutnick A. I., Millman I., Blumberg B. S. Specificities of human antibodies to Australia antigen. Proc Soc Exp Biol Med. 1970 Jun;134(2):548–557. doi: 10.3181/00379727-134-34833. [DOI] [PubMed] [Google Scholar]
  23. Sukeno N., Shirachi R., Yamaguchi J., Ishida N. Reduction and reoxidation of Australia antigen: loss and reconstitution of particle structure and antigenicity. J Virol. 1972 Jan;9(1):182–183. doi: 10.1128/jvi.9.1.182-183.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Summers D. F., Maizel J. V., Jr, Darnell J. E., Jr Evidence for virus-specific noncapsid proteins in poliovirus-infected HeLa cells. Proc Natl Acad Sci U S A. 1965 Aug;54(2):505–513. doi: 10.1073/pnas.54.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. YPHANTIS D. A. EQUILIBRIUM ULTRACENTRIFUGATION OF DILUTE SOLUTIONS. Biochemistry. 1964 Mar;3:297–317. doi: 10.1021/bi00891a003. [DOI] [PubMed] [Google Scholar]
  26. Zuckerman A. J., Baines P. M., Almeida J. D. Australia antigen as a marker of propagation of the serum hepatitis virus in liver cultures. Nature. 1972 Mar 10;236(5341):78–81. doi: 10.1038/236078a0. [DOI] [PMC free article] [PubMed] [Google Scholar]

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