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

Author:

Gerin J. L.1,Holland P. V.2,Purcell R. H.3

Affiliation:

1. Rockville Laboratory of the Molecular Anatomy Program, Oak Ridge National Laboratory, Rockville, Maryland 20852

2. Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20014

3. Blood Bank Department, Clinical Center, National Institutes of Health, Bethesda, Maryland 20014

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.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

Reference23 articles.

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2. Particles associated with Australia antigen in the sera of patients with leukemia, Down's syndrome, and hepatitis;Bayer M.;Nature (London),1968

3. Bishop B. 1966. Digital computation of sedimentation coefficients in zonal centrifuges p. 175-188. In N. G. Anderson (ed.) The development of zonal centrifuges. Nat. Cancer Inst. Monogr. No. 21.

4. A serum antigen (Australia antigen) in Down's syndrome, leukemia, and hepatitis;Blumberg B.;Ann. Intern. Med,1967

5. A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel clectrophoresis;Chrambach A.;Anal. Biochem.,1967

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