Proteins of the Group B Arbovirus Kunjin

Westaway, E. G.; Reedman, Beverley M.

doi:10.1128/jvi.4.5.688-693.1969

Cited by 49 publications

(18 citation statements)

References 18 publications

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“…Structural protein nomenclature. Flaviviruses contain three structural proteins originally designated V-1, V-2, and V-3 (Table 1) by various authors (33,35,39,41,42). Although this nomenclature does provide a general identification, it fails to describe any of the now known chemical, physical, or immunological properties of the proteins.…”

Section: Resultsmentioning

confidence: 99%

“…The major envelope protein of JE and SLE viruses, released from the visions or infected cells after detergent treatment, binds to erythrocytes (17,34), reacts with antibodies in the CF and immunodiffusion tests (13,17,24,42), and induces the formation of neutralizing antibodies (12,26,27). In both CF and immunodiffusion tests, this isolated glycoprotein is serologically broadly cross-reactive (19,26,27).…”

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confidence: 99%

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Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing

Trent¹

1977

J Virol

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Isoelectrofocusing of nonionic-detergent-disrupted flaviviruses separated the envelope glycoprotein of 53,000 to 58,000 daltons and the nucleocapsid protein of 14,000 daltons. The envelope protein and nucleocapsid protein were isolated at isoelectric points of pI 7.8 and 10.3, respectively. The antigenic determinants of St. Louis encephalitis, Japanese encephalitis, and dengue virus envelope and nucleocapsid proteins were examined by solid-phase competition radioimmunoassay. By the appropriate selection of antiserum and competing proteins, it was possible to distinguish type-specific, complex-reactive and flavivirus groupreactive antigenic determinants. The envelope glycoproteins of St. Louis encephalitis, Japanese encephalitis, and dengue viruses were found to contain each of these three classes of antigenic determinants. Most of the determinants on the envelope protein were type specific, some were complex reactive, and a small fraction were flavivirus group reactive. The nucleocapsid protein contained only flavivirus group-reactive antigenic determinants. Purified flaviviruses contain three classes of antigenic determinants: type specific, complex reactive, and flavivirus group reactive. Type-MATERIALS AND METHODS Viruses and cells. Virus strains used in these experiments were the Tampa Bay human-28 strain of SLE and the Nakayama, JaGAr-01, and Yokoshiba strains of JE viruses. Four prototype dengue 608

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing

Trent¹

1977

J Virol

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“…Peak fractions were then pooled and stored at 4". The indirect ELISA to detect purified monoclonal antibody was performed using 96-well U-bottom plates coated with MVE virus purified by the method of Westaway and Reedman (14).…”

Section: Monoclonal Antibody Purificationmentioning

confidence: 99%

An Enzyme Immunoassay to Detect Australian Flaviviruses and Identify the Encephalitic Subgroup using Monoclonal Antibodies

1987

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Summary An antigen‐capture enzyme‐linked immunosorbent assay (ELISA) has been developed to detect antigens of Australian flaviviruses in mosquito pools, suckling mouse brain and infected cell culture supernatant fluid. A monoclonal antibody reactive to an epitope on the envelope glycoprotein common to all flaviviruses was used as the capture antibody. Purified rabbit IgG, produced against Murray Valley encephalitis (MVE) virus, which reacted with eight Australian flaviviruses in haemagglutination inhibition (HI) and in an indirect fluorescent antibody test, was used as the indicator antibody in direct and indirect antigen‐capture ELISA. A monoclonal antibody specific for a subgroup of encephalitic flaviviruses was conjugated to horseradish peroxidase and used as the indicator antibody to distinguish MVE, Kunjin and Alfuy viruses from the remainder tested. This ELISA could detect viral antigen in mosquito cell culture fluids and suckling mouse brain preparations at titres as low as 1000 TCID50/100μ1. Viral antigen in a single mosquito infected with MVE could be detected in a pool of 500.

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“…1. Amino acid labeling and electrophoresis (15) showed that all major proteins are virus-specific. An average recovery of 60% of infectivity was obtained in 18 experiments with JBE, Kunjin, Murray Valley encephalitis, St. Louis encephalitis, and West Nile viruses.…”

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Rapid Preparation of Hemagglutinins of Togaviruses from Infected Cell Culture Fluids

Della-Porta

Westaway

1972

Applied Microbiology

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Antigens in infected cell culture fluids can be easily concentrated by polyethylene glycol precipitation to yield suitable hemagglutinating and complement-fixing antigens for several togaviruses. Although accumulation of hemagglutinin (HA) of Japanese B encephalitis (JBE) virus in infected cell culture fluids has been described (5, 8), little or no HA activity has been recovered for other togaviruses (9, 10). [The term togavirus was recommended by the Subcommittee of Vertebrate Viruses (1) to the International Committee on Nomenclature of Viruses for the arboviruses belonging to serogroups A and B.] We routinely prepare HA of slowgrowing group B and other togaviruses for HAinhibition (HI) tests by concentrating infected culture fluids by using polyethylene glycol (PEG) precipitation (7, 15). Confluent monolayers of PS cells or Vero cells are inoculated with 1/100 infected mouse brain stock for 1 hr, washed, and incubated at 37 C in Eagle's medium or medium 199 supplemented with 0.2% bovine serum albumin. Culture methods, cell lines, and virus strains employed were described previously (6, 13, 14). When cytopathic effect develops, harvested fluids are clarified at 10,000 x g for 10 min at 4 C and then made 8% with respect to PEG 6000 (Koch-Light Ltd., Colnbrook, Bucks, England) by adding 40% (w/v) stock solution at pH 7.8 in Hanks salt solution. After 1 hr at 4 C, virus is sedimented at 10,000 x g for 30 min, the fluid is decanted and drained, and This investigation was supported by a grant from the National Health and Medical Research Council of Australia.

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Proteins of the Group B Arbovirus Kunjin

Cited by 49 publications

References 18 publications

Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing

Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing

An Enzyme Immunoassay to Detect Australian Flaviviruses and Identify the Encephalitic Subgroup using Monoclonal Antibodies

Rapid Preparation of Hemagglutinins of Togaviruses from Infected Cell Culture Fluids

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