1887

Abstract

Summary

Owing to the difficulties of achieving consistently good recoveries of virus from allantoic fluid containing influenza virus by chromatography on calcium phosphate columns the factors determining optimum yield were systematically investigated. It was found that calcium phosphate prepared by precipitation at a controlled pH of 7·5 gave best results provided that eluting buffers were adjusted to the same pH. Although calcium phosphate fractions were still evidently contaminated with non-viral material, further purification could be obtained by chromatography in agarose-gel columns. It appeared that separation of the virus on these columns was probably achieved as a result of the binding of virus surface groups on to the gel. A minimum overall purification factor of about 800-fold was achieved.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-1-1-49
1967-01-01
2022-01-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/1/1/JV0010010049.html?itemId=/content/journal/jgv/10.1099/0022-1317-1-1-49&mimeType=html&fmt=ahah

References

  1. Ackers G. K., Steere R. L. 1962; Restricted diffusion of macromolecules through agar-gel membranes. Biochim. biophys. Acta 59:137
    [Google Scholar]
  2. Bengtsson S., Pffllipson L. 1964; Chromatography of animal viruses on pearl-condensed agar. Biochim. biophys. Acta 79:399
    [Google Scholar]
  3. Bernardi G. 1965; Chromatography of nucleic acids on hydroxyapatite. Nature; Lond: 206779
    [Google Scholar]
  4. Biddle F., Pepper D. S., Belyavin G. 1965; Properties of horse serum gamma inhibitor. Nature; Lond: 207381
    [Google Scholar]
  5. Cech M. 1962; Some notes on the use of agar gel filtration in the study of plant viruses. Virology 18:487
    [Google Scholar]
  6. Choppin P. W., Tamm I. 1960; Studies of two kinds of virus particle which comprise influenza A-2 virus strains. J. exp. Med 112:895
    [Google Scholar]
  7. Curtain C. C. 1954; The adsorption of influenza virus by haemagglutination inhibitors coupled to powdered cellulose. Br. J. exp. Path 35:255
    [Google Scholar]
  8. Hjerten S. 1962; Chromatographic separation according to size of macromolecules and cell particles on columns of agarose suspension. Archs Biochem. Biophys 99:466
    [Google Scholar]
  9. Hjerten S. 1964; The preparation of agarose spheres for chromatography of molecules and particles. Biochim. biophys. Acta 79:393
    [Google Scholar]
  10. Kibardin S. A. 1965; The chromatography of biopolymers on calcium phosphate gels. Russ. chem. Revs 34:629
    [Google Scholar]
  11. Main R. K., Wilkins M. J., Cole L. J. 1959; Modified calcium phosphate for column chromatography of polynucleotides and proteins. J. Am. chem. Soc 81:6490
    [Google Scholar]
  12. Miles A. A., Misra S. S. 1938; The estimation of the bactericidal power of blood. J. Hyg., Camb 38:732
    [Google Scholar]
  13. Poison A. 1961; Fractionation of protein mixtures on columns of granulated agar. Biochim. biophys. Acta 50:565
    [Google Scholar]
  14. Siegel L. M., Monty K. J. 1966; Determination of the molecular parameters of impure proteins by combined gel filtration and density gradient centrifugation. Biochim. biophys. Acta 112:346
    [Google Scholar]
  15. Squire P. G. 1964; A relationship between the molecular weight of macromolecules and their elution volumes based on a model for Sephadex gel filtration. Archs Biochem. Biophys 107:471
    [Google Scholar]
  16. Steere R. L., Ackers G. K. 1962a; Purification and separation of tobacco mosaic virus and southern bean mosaic virus by agar-gel filtration. Nature; Lond: 194114
    [Google Scholar]
  17. Steere R. L., Ackers G. K. 1962b; Restricted diffusion chromatography through calibrated columns of granulated agar gel; a simple method of particle-size determination. Nature; Lond: 196475
    [Google Scholar]
  18. Steere R. L. 1963; Tobacco mosaic virus: purifying and sorting associated particles according to length. Science; N.Y: 1401089
    [Google Scholar]
  19. Taverne J., Marshall J. H., Fulton F. 1958; The purification and concentration of viruses and virus soluble antigens on calcium phosphate. J. gen. Microbiol 19:451
    [Google Scholar]
  20. Tiselius A., Hjerten S., Levin O. 1956; Protein chromatography on calcium phosphate columns. Archs Biochem. Biophys 65:132
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-1-1-49
Loading
/content/journal/jgv/10.1099/0022-1317-1-1-49
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error