FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Clin Microbiol / Chromatographic separations of alphavirus strains by hydroxy...
Journal of clinical microbiology1977; 6(3); 238-243; doi: 10.1128/jcm.6.3.238-243.1977

Chromatographic separations of alphavirus strains by hydroxylapatite.

Abstract: Hydroxylapatite column chromatography methods were developed to characterize selected alphavirus populations. Different conditions of pH and phosphate molarity were required to obtain satisfactory elution profiles and separations for Western equine encephalomyelitis virus strains, compared with Eastern equine encephalomyelitis virus and Semliki Forest virus strains. Raising the pH of the buffers effected earlier elutions of all viruses. Selection of phosphate gradients with more gentle slopes and adjustment to the proper pH effected better separations of virus subpopulations. Elution profiles were not affected by 0.85% NaCl, 10% fetal calf serum, or 1% bovine serum albumin, which are common constituents of virus-stabilizing diluents. Passage of Western equine encephalomyelitis and Semliki Forest viruses in BHK-21, Vero, or duck embryo cell cultures or in suckling mouse brains did not usually affect elution profiles, unless passage also resulted in a shift in the plaque size marker. Essentially all infectious virus applied to the column was recoverable in appropriate fractions. This permitted accurate determinations of heterogeneity within alphavirus populations. For Western equine encephalomyelitis large-plaque (LP) and small-plaque (SP) virus populations, it was possible to detect ratios of 1 LP in a population of 10(6) SP, and 1 SP in 10(3) LP by using linear phosphate gradients. When stepwise elution procedures were used, it was possible to detect ratios of 1 SP in a population of 10(5) LP. Hydroxylapatite column chromatography therefore appears to be a useful tool for characterizing alphaviruses and for isolating minority subpopulations of viruses of biological or epidemiological importance from apparently homogeneous virus stocks.
Get Full Text
Publication Date: 1977-09-01 PubMed ID: 20452PubMed Central: PMC274746DOI: 10.1128/jcm.6.3.238-243.1977Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article is about how hydroxylapatite column chromatography can be used to separate different strains of alphavirus and detect minor subpopulations within those strains. The research also indicates that this method of chromatography could be a significant tool for characterizing alphaviruses.

Hydroxylapatite Column Chromatography Application

  • Methods of hydroxylapatite column chromatography were developed specifically to analyze separate alphavirus populations.
  • Various conditions of pH and phosphate molarity were required to achieve satisfactory separation and elution profiles for Western equine encephalomyelitis virus strains as opposed to Eastern equine encephalomyelitis virus and Semliki Forest virus strains.
  • Increasing the pH of the buffers resulted in earlier elutions of all viruses.
  • Better separation of virus subpopulations was achieved by selecting phosphate gradients with gentler slopes and adjusting to the right pH.

Chromatography Performance with Other Components

  • Elution profiles were unaffected by substances such as 0.85% NaCl, 10% fetal calf serum, or 1% bovine serum albumin, which are typically present in virus-stabilizing diluents.
  • The passage of Western equine encephalomyelitis and Semliki Forest viruses in BHK-21, Vero, or duck embryo cell cultures or in suckling mouse brains did not usually impact elution profiles unless the passage also resulted in a shift in the plaque size marker.

Assessment of Alphavirus Populations

  • Almost all infectious viruses applied to the column could be recovered in relevant fractions. This enabled accurate determination of heterogeneity within alphavirus populations.
  • For Western equine encephalomyelitis large-plaque (LP) and small-plaque (SP) virus populations, the presence of 1 LP in a population of 10(6) SP and 1 SP in 10(3) LP was detectable by using linear phosphate gradients.
  • When stepwise elution procedures were applied, it was possible to pinpoint ratios of 1 SP in a population of 10(5) LP.

Implications of the Research

  • Hydroxylapatite column chromatography is presented as a valuable tool for the characterization of alphaviruses.
  • The technique allows for the isolation of minor, potentially important subpopulations of viruses from seemingly homogeneous virus stocks.

Cite This Article

APA
Jahrling PB, Beall JL. (1977). Chromatographic separations of alphavirus strains by hydroxylapatite. J Clin Microbiol, 6(3), 238-243. https://doi.org/10.1128/jcm.6.3.238-243.1977

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 6
Issue: 3
Pages: 238-243

Researcher Affiliations

Jahrling, P B
    Beall, J L

      MeSH Terms

      • Buffers
      • Chromatography / methods
      • Encephalitis Virus, Western Equine / isolation & purification
      • Hydrogen-Ion Concentration
      • Hydroxyapatites
      • Phosphates
      • Semliki forest virus / isolation & purification

      References

      This article includes 9 references
      1. Jahrling PB, Gorelkin L. Selective clearance of a benign clone of Venezuelan equine encephalitis virus from hamster plasma by hepatic reticuloendothelial cells.. J Infect Dis 1975 Dec;132(6):667-76.
        pubmed: 1202111doi: 10.1093/infdis/132.6.667google scholar: lookup
      2. OZAKI Y, DIWAN AR, TAKIZAWA M, MELNICK JL. CHROMATOGRAPHY OF POLIOVIRUS ON CALCIUM PHOSPHATE AND ITS APPLICATION TO THE IDENTIFICATION OF VACCINE PROGENY STRAINS.. J Bacteriol 1965 Mar;89(3):603-10.
        pubmed: 14273634doi: 10.1128/jb.89.3.603-610.1965google scholar: lookup
      3. Burness AT. Separation of plaque-type variants of encephalomyocarditis virus by chromatography on calcium phosphate.. J Virol 1967 Apr;1(2):308-16.
        pubmed: 4318950doi: 10.1128/JVI.1.2.308-316.1967google scholar: lookup
      4. Fleming P. Semliki forest virus-chick embryo cell interactions.. J Gen Virol 1973 Jun;19(3):353-67.
        pubmed: 4723534doi: 10.1099/0022-1317-19-3-353google scholar: lookup
      5. Jahrling PB, Dendy E, Eddy GA. Correlates to increased lethality of attenuated Venezuelan encephalitis virus vaccine for immunosuppressed hamsters.. Infect Immun 1974 May;9(5):924-30.
        pubmed: 4824635doi: 10.1128/iai.9.5.924-930.1974google scholar: lookup
      6. Pedersen CE Jr, Slocum DR, Levitt NH. Chromatography of Venezuelan equine encephalomyelitis virus strains on calcium phosphate.. Appl Microbiol 1972 Jul;24(1):91-5.
        pubmed: 5057376doi: 10.1128/am.24.1.91-95.1972google scholar: lookup
      7. Bradish CJ, Allner K, Maber HB. The virulence of original and derived strains of Semliki forest virus for mice, guinea-pigs and rabbits.. J Gen Virol 1971 Aug;12(2):141-60.
        pubmed: 5123325doi: 10.1099/0022-1317-12-2-141google scholar: lookup
      8. Young NA, Johnson KM. Antigenic variants of Venezuelan equine encephalitis virus: their geographic distribution and epidemiologic significance.. Am J Epidemiol 1969 Mar;89(3):286-307.
        pubmed: 5773424doi: 10.1093/oxfordjournals.aje.a120942google scholar: lookup
      9. Jahrling PB. Virulence heterogeneity of a predominantly avirulent Western equine encephalitis virus population.. J Gen Virol 1976 Jul;32(1):121-8.
        pubmed: 956785doi: 10.1099/0022-1317-32-1-121google scholar: lookup

      Citations

      This article has been cited 4 times.
      1. Byrnes AP, Griffin DE. Binding of Sindbis virus to cell surface heparan sulfate. J Virol 1998 Sep;72(9):7349-56.
        doi: 10.1128/JVI.72.9.7349-7356.1998pubmed: 9696831google scholar: lookup
      2. Jahrling PB, Hesse RA, Metzger JF. Radioimmunoassay for quantitation of antibodies to alphaviruses with staphylococcal protein A. J Clin Microbiol 1978 Jul;8(1):54-60.
        doi: 10.1128/jcm.8.1.54-60.1978pubmed: 566768google scholar: lookup
      3. Osborne LC, Georgiades JA, Johnson HM. Large-scale production and partial purification of mouse immune interferon. Infect Immun 1979 Jan;23(1):80-6.
        doi: 10.1128/iai.23.1.80-86.1979pubmed: 422237google scholar: lookup
      4. Brandt WE, McCown JM, Top FH Jr, Bancroft WH, Russell PK. Effect of passage history on dengue-2 virus replication in subpopulations of human leukocytes. Infect Immun 1979 Nov;26(2):534-41.
        doi: 10.1128/iai.26.2.534-541.1979pubmed: 121109google scholar: lookup
      Subscribe to our newsletter
      Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.