Summary
The efficiency of differential centrifugation, isopycnic banding, anionic exchange chromatography and gelfiltration have been assayed with respect to capacity to separate adenovirus type 4 components. These results are presented and the following procedure resulting in separation of six different viral components, all of which have been identified in the electron microscope in their purified state, is proposed.
The first stage, which comprised differential centrifugation in a discontinuous CsCl gradient of crude virus material, was followed by repeated isopycnic banding of the “non-soluble” components thus resulting in purified intact virus and empty capsids.
In the second stage all the soluble components were at least partially separated by anionic exchange chromatography. They eluted in the following sequence: fibers, hexons, pentons and soluble complete HA.
In the subsequent stage zonal centrifugation was applied to obtain purified hexons and soluble complete HA, whereas application of gelfiltration was needed to purify pentons from contaminating hexons.
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References
Anderson, N. G.: Zonal centrifuges and other separation systems. Science154, 103–112 (1966).
Boyer, G. S., C. Leuchtenberger, andH. S. Ginsberg: Cytological and cytochemical studies of HeLa cells infected with, adenoviruses. J. exp. Med.105, 195–216 (1957).
Denny, F. W., Jr., andH. S. Ginsberg: Certain biological characteristics of adenoviruses, types 5, 6, 7 and 14. J. Immunol.86, 567–574 (1961).
Gelderblom, H., R. Wigand, andH. Bauer: DEAE-chromatography of adenoviruses of Rosen's group II. Nature (Lond.)205, 625–626 (1965).
Ginsberg, H. S.: Characteristics of the new respiratory viruses (adenoviruses). I. Qualitative and quantitative aspects of the neutralization reaction. J. Immunol.77, 271–278 (1956).
Ginsberg, H. S., H. G. Pereira, R. C. Valentine, andW. C. Wilcox: A proposed terminology for the adenovirus antigens and virion morphological subunits. Virology28, 782–783 (1966).
Handbook of chemistry and physics, 1966–1967. CRC47, C-706.
Hilleman, M. R., A. J. Tousimis, andJ. H. Werner: Biophysical characterization of the RI (RI-67) viruses. Proc. Soc. exp. Biol. (N. Y.)89, 587–593 (1955).
Haruna, J., H. Yaoi, R. Kono, andI. Watanabe: Separation of adenoviruses by chromatography on DEAE-cellulose. Virology13, 264–267 (1961).
Hu, A. S. L., R. M. Bock, andH. O. Halvorson: Separation of labeled from unlabeled proteins by equilibrium density gradient sedimentation. Ann. Biochem.4, 489–504 (1962).
Huebner, R. J., W. P. Rowe, andR. M. Chanock: Newly recognized respiratory tract viruses. Ann. Rev. Microbiol.12, 49–76 (1958).
Huebner, R. J., H. G. Pereira, A. C. Allison, A. C. Hollinshead, andH. C. Turner: Production of type specific C antigen in virus-free bamster tumor cells induced by adenovirus type 12. Proc. nat. Acad. Sci. (Wash.)51, 432–439 (1964).
Huebner, R. J., M. J. Chasey, R. M. Chanock, andK. Schell: Tumors induced in hamsters by a strain of adenovirus type 3: Sharing of tumor-antigens and neoantigens with tbose produced by adenovirus type 7 tumor. Proc. nat. Acad. Sci. (Wash.)54, 381–388 (1965).
Kasel, J. A., andM. Huber: Relationship of the adenovirus erythrocytereceptor-modifying factor to the type specific complement fixing antigen. Proc. Soc. exp. Biol. (N. Y.)116, 16–18 (1964).
Klemperer, H. G., andH. G. Pereira: Study of adenovirus antigens fractionated by chromatography on DEAE-cellulose. Virology9, 536–545 (1959).
Köhler, K.: Reinigung und Charakterisierung zweier Proteine des Adenovirus Typ 2. Z. Naturforsch.20b, 747–752 (1965).
Lacy, S., andM. Green: Adenovirus multiplication: Genetic relatedness of tumorigenic human adenovirus types 7, 12 and 18. Science150, 1296–1298 (1965).
Laurent, T. C., andJ. Killander: A theory of gelfiltration and its experimental verification. J. Chromatogr.14, 317–330 (1964).
Meselson, M., F. W. Stahl, andJ. Vinograd: Equilibrium sedimentation of macromolecules in density gradients. Proc. nat. Acad. Sci. (Wash.)43, 581–588 (1957).
Norrby, E.: The relationship between the soluble antigens and the virion of adenovirus type 3. I. Morphological characteristics. Virology28, 236–248 (1966a).
Norrby, E.: The relationship between the soluble antigens and the virion of adenovirus type 3. II. Identification and characterization of an incomplete hemagglutinin. Virology30, 608–617 (1966b).
Norrby, E., M. Grönberg, andP. Magnusson: Virus-specific products appearing during the multiplication of adenovirus type 3. Unio Nordica Contra Cancrum, Symp. Virus and Cancer, pp. 118–124 (1964).
Norrby, E., andP. Skaaret: The relationship between soluble antigens and the virion of adenovirus type 3. III. Immunological identification of fiber antigen and isolated vertex capsomer antigen. Submitted for publication to Virology (1967).
Norrby, E., andG. Wadell: Soluble components of adenovirustype 4. Virology31, 592–600 (1967).
Pereira, H. G., andM. V. T. de Figueiredo: Mechanism of hemagglutination by adenovirus types 1, 2, 4, 5 and 6. Virology18, 1–8 (1962).
Philipson, L.: Separation on DEAE cellulose of components associated with adenovirus reproduction. Virology10, 459–465 (1960).
Rosen, L.: A hemagglutination-inhibition technique for typing adenovirus. Amer. J. Hyg.71, 120–128 (1960).
Sever, J. L.: Application of a microtechnique to viral serological investigations. J. Immunol.88, 320–329 (1961).
Siegel, L. M., andK. J. Monty: Determination of molecular weights and fractional ratios of proteins in impure systems by use of gelfiltration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim. biophys. Acta (Amst.)112, 346–362 (1966).
Simon, M.: Chromatography of adenoviruses on calcium phosphate columns. Acta virol.6, 302–308 (1962).
Sober, H. A., andE. A. Peterson: Protein chromatography on ion exchange cellulose. Fed. Proc.117, 1116–1126 (1958).
Wadell, G., E. Norrby, andU. Schönning: Ultrastructure of soluble antigens and the virions of adenovirus type 4. Arch. ges. Virusforsch.21, 234 (1967).
Wilcox, W. C., andH. S. Ginsberg: Purification and immunological characterization of types 4 and 5 adenovirus-soluble antigens. Proc. nat. Acad. Sci. (Wash.)47, 512–526 (1961).
Wilcox, W. C., andH. S. Ginsberg: Structure of type 5 adenovirus. I. Antigenie relationship of virus-structural proteins to virus-specific soluble antigens from infected cells. J. exp. Med.118, 295–306 (1963).
Zhdanov, V. M., andL. B. Mekler: Correlation between the surface structure of viruses and latency. Nature (Lond.)195, 924 (1962).
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Supported by grants from the Swedish Medical Research Council (Project No. K67-16X-548-03).
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Wadell, G., Skaaret, P. Separation of adenovirus type 4 components. Archiv f Virusforschung 21, 366–382 (1967). https://doi.org/10.1007/BF01241737
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DOI: https://doi.org/10.1007/BF01241737