The use of single-radial-haemolysis for rubella antibody studies

Clarke, M.; Boustred, Janet; Seagroatt, Valerie; Schild, G. C.

doi:10.1017/s0022172400053195

Cited by 24 publications

(10 citation statements)

References 8 publications

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“…This has been noted with influenza viruses (Schild et al, 1975;RusseU et al, 1975;Oxford et al, 1981;Farrohi et al, 1977) and other viruses including mumps (Norrby et al, 1977;V~i~n~inen et al, 1976) and rubella (Clarke et al, 1977;Gaidamovich et al, 1980). Several of these authors commented on the ability to measure lower levels of antibody by SRH than in the HI test because the sera did not have to be treated to remove non-specific inhibitors of haemagglutination.…”

Section: Antibody Responses To Vaccinesmentioning

confidence: 80%

Antibody and Cytotoxic T Lymphocyte Responses of Humans to Live and Inactivated Influenza Vaccines

Ennis

Schild

1982

Journal of General Virology

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SUMMARYThe antibody and HLA-restricted virus-specific cytotoxic T lymphocyte (CTL) responses to influenza vaccine of 36 volunteers were analysed. Three vaccines were used: a live attenuated, and two types of inactivated, a whole virus and a purified surface antigen vaccine. Antibody to haemagglutinin (HA) was assayed using plaque neutralization, single-radial-haemolysis and haemagglutination-inhibition (HI) techniques. Antibodies to nucleoprotein and matrix antigens were also measured. Most of the volunteers had antibody responses to the HA in the inactivated vaccines which were detected by all three techniques. Nine of the twelve recipients of the live virus vaccine did not have an antibody response detected by the HI test, but four of these did have antibody responses when the plaque neutralization test was used. Single-radial-haemolysis was more sensitive than the HI test for detecting low levels of antibody, but the plaque neutralization test was the most sensitive for detecting low levels of antibody. Most volunteers had a rise in their HLA-restricted influenza-specific memory CTL response, but three recipients of live vaccine who did not have an antibody response by any technique also did not have an increase in their cytotoxic T cell activity. Three volunteers, two of whom had received live vaccine, had a positive CTL response without antibody response.

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Section: Antibody Responses To Vaccinesmentioning

confidence: 80%

Antibody and Cytotoxic T Lymphocyte Responses of Humans to Live and Inactivated Influenza Vaccines

Ennis

Schild

1982

Journal of General Virology

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“…This assay was carried out using a modification of the method described by Clarke et al [8]. Briefly, sheep red blood cells sensitirctl with rubella antigen were reacted with test samples in the presence of complement in an agarose gel.…”

Section: Single Rudial Haemolysismentioning

confidence: 99%

“…One product (WIG B) was not degraded by either plasmin or kallikrein treatments (figs. 7,8). This product is formulated at an acidic pH of 4.25; all other samples are buffered between a pH range of 6.4-7.2.…”

Section: Ejfects Ofph On Proteolytic Susceptibilitymentioning

confidence: 99%

Fragmentation of Therapeutic Human Immunoglobulin Preparations

et al. 1995

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Some commercial batches of human therapeutic immunoglobulins (Ig) have been found to show evidence of molecular fragmentation when examined by molecular sizing methodologies including sodium dodecyl sulphate polyacrylamide gel electrophoresis [SDS-PAGE] and size exclusion high performance liquid chromatography (SE-HPLC). These batches all demonstrated impaired immunobiological activity (efficacy) as assessed by Fc function measured using a rubella haemolytic assay and as such are likely to be subpotent for therapeutic use. Fragmented Igs were characterized by the presence of at least three protein bands and peaks additional to monomeric IgG. Incubation of Igs with blood enzymes (plasmin and kallikrein) reproduced the fragmentation patterns observed for intrinsically degraded batches, suggesting that fragmentation occurred by contamination with these proteases from the source material (human blood) during manufacture. Intravenous Igs (IVIG) were found to be more susceptible to proteolysis than intramuscular Igs, probably as a consequence of the post-fractionation processing that some IVIGs receive which may induce molecular alterations, allowing enzyme access and fragmentation. Two of the products examined were found to be relatively resistant to proteolysis and both were formulated by processes that limit enzyme activity. These processes were inclusion of an enzyme inhibitor, alpha 2-macroglobulin, and formulation at acidic pH. Enzyme carry-over into the final product is a likely cause of Ig fragmentation, and reduction in levels of such contamination should lead to improvements in product stability and efficacy.

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“…The main advantage is the ability to simultaneously and rapidly test a large number of samples without pre-treatment (excluding complement inactivation) and the requirement for only a small volume of serum samples. These features make the SRH assay a particularly suitable assay for large-scale investigation especially for epidemiological studies (Clarke et al, 1977;Callow and Beare, 1976;Trombetta et al, 2014).…”

Section: Introductionmentioning

confidence: 99%