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Biology subjects

Scott, S. D.

Publications and source records attributed to Scott, S. D..

2 recordsLinked to original sources

Application and comparison of lyophilisation protocols to enhance stable long-term storage of filovirus pseudotypes for use in antibody neutralisation tests

Filoviruses encompass highly pathogenic viruses placing sporadic public health burden on countries affected. Efforts for improved diagnostics and surveillance are needed considering the recent Ebola outbreaks in Africa. The need for high containment facilities can be circumvented by the use of pseudotype viruses (PV), which can be handled in low containment, for tropism, drug screening, vaccine immunogenicity and serosurveillance studies. In this study we assessed stability and functionality after long-term storage of lyophilised filovirus pseudotypes for use in neutralisation assays. Lyophilised Ebola and Marburg PVs retained production titres for at least two years when stored at +4{degrees}C or less. Lyophilised Ebola PVs performed similarly to non-lyophilised PVs in neutralisation assays after reconstitution. When stored at high temperatures (+37{degrees}C), lyophilised PVs did not retain titres after one-month storage, however, when lyophilised using pilot scale facilities EBOV PVs retained titres and performed well in neutralisation assays after one-month storage at 37{degrees}C suggesting removing residual moisture might be crucial for avoiding cold-chain transportation. Lyophilisation could allow reagents to be transported more efficiently as well as reducing costs for a future serological kit.

microbiology↗

Establishment of pan-Influenza A (H1-H18) and pan-Influenza B (pre-split, Vic/Yam) Pseudotype Libraries for efficient vaccine antigen selection

We have developed an influenza hemagglutinin (HA) pseudotype library encompassing Influenza A subtypes HA1-18, and Influenza B subtypes (both lineages) to be employed in influenza pseudotype microneutralization (pMN) assays. The pMN is highly sensitive and specific for detecting virus-specific neutralizing antibodies against influenza viruses and can be used to assess antibody functionality in vitro. Here we show the production of these viral HA pseudotypes and their employment as substitutes for wildtype viruses in influenza serological and neutralization assays. We demonstrate its utility in detecting serum response to vaccination with the ability to evaluate cross-subtype neutralizing responses elicited by specific vaccinating antigens. Our findings may inform further pre-clinical studies involving immunization dosing regimens in mice and may help in the creation and selection of better antigens for vaccine design. These HA pseudotypes can be harnessed to meet strategic objectives that contribute to the strengthening of global influenza surveillance, expansion of seasonal influenza prevention and control policies, and strengthening pandemic preparedness and response.

immunology↗