Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1971 May;7(5):569–576. doi: 10.1128/jvi.7.5.569-576.1971

Australia Antigen: Large-Scale Purification from Human Serum and Biochemical Studies of Its Proteins

J L Gerin a, P V Holland b, R H Purcell c
PMCID: PMC356164  PMID: 5105002

Abstract

Biophysical techniques are described for the large-scale isolation of Australia antigen (Au) from unit quantities of human serum by using the batch-type zonal centrifuge rotors. A three-step procedure involving isopycnic banding of the particle in CsCl density gradients and rate-zonal centrifugation on sucrose gradients resulted in a highly purified Au preparation which was used for biochemical studies of Au proteins and as immunizing antigen for the production of reagent antiserum in animals. The spherical form of Au, which was devoid of detectable nucleic acid, was composed of two major proteins (AuP1 and AuP2) and a minor protein (AuP3) of 26,000, 32,000, and 40,000 molecular weight, respectively, as determined by acrylamide gel electrophoresis. The significance of these findings to the possibility of Au subtypes is discussed.

Full text

PDF
573

Selected References

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

  1. Anderson N. G., Nunley C. E., Rankin C. T., Jr Analytical techniques for cell fractions. XV. Rotor B-XXIX--a new high-resolution zonal centrifuge rotor for virus isolation and cell fractionation. Anal Biochem. 1969 Oct 1;31(1):255–271. doi: 10.1016/0003-2697(69)90265-6. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bishop B. S. Digital computation of sedimentation coefficients in zonal centrifuges. Natl Cancer Inst Monogr. 1966 Jun;21:175–188. [PubMed] [Google Scholar]
  4. Blumberg B. S., Gerstley B. J., Hungerford D. A., London W. T., Sutnick A. I. A serum antigen (Australia antigen) in Down's syndrome, leukemia, and hepatitis. Ann Intern Med. 1967 May;66(5):924–931. doi: 10.7326/0003-4819-66-5-924. [DOI] [PubMed] [Google Scholar]
  5. Chrambach A., Reisfeld R. A., Wyckoff M., Zaccari J. A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Anal Biochem. 1967 Jul;20(1):150–154. doi: 10.1016/0003-2697(67)90272-2. [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. Gerin J. L., Purcell R. H., Hoggan M. D., Holland P. V., Chanock R. M. Biophysical properties of Australia antigen. J Virol. 1969 Nov;4(5):763–768. doi: 10.1128/jvi.4.5.763-768.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giles J. P., McCollum R. W., Berndtson L. W., Jr, Krugman S. Relation of Australia-SH antigen to the willowbrook MS-2 strain. N Engl J Med. 1969 Jul 17;281(3):119–122. doi: 10.1056/NEJM196907172810302. [DOI] [PubMed] [Google Scholar]
  9. Gocke D. J., Kavey N. B. Hepatitis antigen. Correlation with disease and infectivity of blood-donors. Lancet. 1969 May 31;1(7605):1055–1059. doi: 10.1016/s0140-6736(69)91701-2. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  12. Okochi K., Murakami S. Observations on Australia antigen in Japanese. Vox Sang. 1968;15(5):374–385. doi: 10.1111/j.1423-0410.1968.tb04078.x. [DOI] [PubMed] [Google Scholar]
  13. Pesendorfer F., Krassnitzky O., Wewalka F. Immunelektrophoretischer Nachweis von "Hepatitis-Associated-Antigen" (Au-SH-Antigen. Klin Wochenschr. 1970 Jan 1;48(1):58–59. doi: 10.1007/BF01486135. [DOI] [PubMed] [Google Scholar]
  14. Prince A. M. An antigen detected in the blood during the incubation period of serum hepatitis. Proc Natl Acad Sci U S A. 1968 Jul;60(3):814–821. doi: 10.1073/pnas.60.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Purcell R. H., Gerin J. L., Holland P. V., Cline W. L., Chanock R. M. Preparation and characterization of complement-fixing hepatitis-associated antigen and antiserum. J Infect Dis. 1970 Feb;121(2):222–226. doi: 10.1093/infdis/121.2.222. [DOI] [PubMed] [Google Scholar]
  16. Purcell R. H., Holland P. V., Walsh J. H., Wong D. C., Morrow A. G., Chanock R. M. A complement-fixation test for measuring Australia antigen and antibody. J Infect Dis. 1969 Sep;120(3):383–386. doi: 10.1093/infdis/120.3.383. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Shulman N. R., Barker L. F. Virus-like antigen, antibody, and antigen-antibody complexes in hepatitis measured by complement fixation. Science. 1969 Jul 18;165(3890):304–306. doi: 10.1126/science.165.3890.304. [DOI] [PubMed] [Google Scholar]
  19. Vyas G. N., Shulman N. R. Hemagglutination assay for antigen and antibody associated with viral hepatitis. Science. 1970 Oct 16;170(3955):332–333. doi: 10.1126/science.170.3955.332. [DOI] [PubMed] [Google Scholar]
  20. Walsh J. H., Yalow R., Berson S. A. Detection of Australia antigen and antibody by means of radioimmunoassay techniques. J Infect Dis. 1970 May;121(5):550–554. doi: 10.1093/infdis/121.5.550. [DOI] [PubMed] [Google Scholar]
  21. Wright R. R., Pappas W. S., Carter J. A., Weber C. W. Preparation and recovery of cesium compounds for density gradient solutions. Natl Cancer Inst Monogr. 1966 Jun;21:241–249. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES