1887

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Volume 19, Issue 3

The Purification and Concentration of Viruses and Virus Soluble Antigens on Calcium Phosphate Free

Abstract

SUMMARY: Chromatography on columns of calcium phosphate has been used to purify and concentrate viruses and their related soluble antigens. The method is an adaptation of one which has already proved successful with proteins and other large molecules. An important advantage is that all operations are carried out at pH 7, elution being effected by alterations in phosphate concentration. The PR 8 strain of influenza was used as a model. Chromatrography on a calcium phosphate column effected a 30- to 100-fold increase in purity, with a recovery of 50–80 %. By chromatography on a second small column a 10- to 30-fold concentration could be obtained; little further purification occurred. Material with a purity up to 10 haemaggluti-nation units/mg. protein was obtained; this compares favourably with material purified by other methods. PR 8 soluble antigen was similarly purified and concentrated, and Semliki forest virus was also purified. In preliminary experiments, purification of vaccinia, encephalomyocarditis, Coxsackie and poliomyelitis type III viruses has also been effected. The method is simple and rapid and can be used with crude virus suspensions from different sources. The viruses retained both their infectivity and their antigenicity as judged by complement fixation.

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© Society for Gerenal Microbiology, 1958
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1958-12-01
2023-03-21
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References

  1. Ada G.L., Perry B.T. 1956; Influenza virus nucleic acid: relationship between biological characteristics of the virus particle and properties of the nucleic acid. J. gen. Microbiol 14:623
     [Google Scholar]
  2. Burke D.C., Isaacs A., Walker J. 1957; The nucleic acid content of influenza virus. Biochim. biophys. Acta 26:576
     [Google Scholar]
  3. Elford W.J., Chu C.M., Dawson I.M., Dudgeon J.A., Fulton F., Smiles J. 1948; Physical properties of the viruses of Newcastle disease, fowl plague and mumps. Brit. J. exp. Path 29:590
     [Google Scholar]
  4. Fulton F. 1958; The measurement of complement fixation by viruses. Advanc. Virus Res 5:247
     [Google Scholar]
  5. Fulton F., Armitage P. 1951; Surviving tissue suspensions for influenza virus titration. J. Hyg., Camb 49:247
     [Google Scholar]
  6. Fulton F., Dumbell K.R. 1949; The serological comparison of strains of influenza virus. J. gen. Microbiol 3:97
     [Google Scholar]
  7. Fulton F., Isaacs A. 1953; Influenza virus multiplication in the chick chorioallantoic membrane. J. gen. Microbiol 9:119
     [Google Scholar]
  8. Johnson M.J. 1941; Isolation and properties of a pure yeast polypeptidase. J. biol. Chem 137:575
     [Google Scholar]
  9. Lowry O.H., Rosebrough N.J., Farr A.L., Randall R.J. 1951; Protein measurement with the Folin phenol reagent. J. biol. Chem 193:265
     [Google Scholar]
  10. Main R.K., Cole L.J. 1957; Chromatography of deoxyribonucleic acids on calcium phosphate columns. Arch. Biochem. Biophys 68:186
     [Google Scholar]
  11. Miller H.K., Schlesinger R.W. 1955; Differentiation and purification of influenza viruses by adsorption on aluminium phosphate. J. Immunol 75:155
     [Google Scholar]
  12. Partridge S.M., Westall R.G. 1949; Displacement chromatography on synthetic ion-exchange resins. I. Separation of organic bases and amino acids using cation-exchange resins. Biochem. J 44:418
     [Google Scholar]
  13. Salk J.E. 1941; Partial purification of the virus of epidemic influenza by adsorption on calcium phosphate. Proc. Soc. exp. Biol., N.Y 46:709
     [Google Scholar]
  14. Semenza G. 1957; Chromatographic purification of cysteinylglycinase. Biochim. biophys. Acta 24:401
     [Google Scholar]
  15. Stanley W.M. 1945; The precipitation of purified concentrated influenza virus and vaccine on calcium phosphate. Science 101:332
     [Google Scholar]
  16. Steelman S.L. 1958; Chromatography of follicle-stimulating hormone (FSH) on hydroxylapatite. Biochim. biophys. Acta 27:405
     [Google Scholar]
  17. Tiselius A. 1954; Chromatography of proteins on calcium phosphate columns. Ark. Kemi 7:443
     [Google Scholar]
  18. Tiselius A. 1955; Chromatographic experiments with proteins. Ann. Acad. Sci. fenn A II, 60:257
     [Google Scholar]
  19. Tiselius A., Hjertén S., Levin Ö. 1956; Protein chromatography on calcium phosphate columns. Arch. Biochem. Biophys 65:132
     [Google Scholar]
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The Purification and Concentration of Viruses and Virus Soluble Antigens on Calcium Phosphate
Microbiology 19, 451 (1958); https://doi.org/10.1099/00221287-19-3-451
/content/journal/micro/10.1099/00221287-19-3-451
/content/journal/micro/10.1099/00221287-19-3-451
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