Evaluation of a DAS-ELISA for quantification of foot-and-mouth disease virus 146S antigen during vaccine preparation

dc.contributor.advisorQuan, Melvyn
dc.contributor.coadvisorHeath, Livio Edward
dc.contributor.emailTlakak@arc.agric.zaen_US
dc.contributor.postgraduateTlaka, Kabelo
dc.date.accessioned2024-02-08T11:19:47Z
dc.date.available2024-02-08T11:19:47Z
dc.date.created2024-04-15
dc.date.issued2023-10-01
dc.descriptionMini Dissertation (MSc (Tropical Animal Health))--University of Pretoria, 2023.en_US
dc.description.abstractThe inactivated foot-and-mouth disease (FMD) vaccine's protection is dependent on the intact component of the FMD virus (FMDV) antigen, the 146S antigen particle. Sucrose density gradient (SDG) centrifugation is the standardised method for quantifying 146S antigen during FMD vaccine formulation. However, because it is operator-dependent, this approach is labour-intensive and produces varied outcomes. As a result, the polyclonal double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed as an alternative approach to quantify the intact 146S antigen of both FMDV South African Territories (Mahapatra and Parida) -1 & 2 serotypes. The polyclonal DAS-ELISA performance was compared to the SDG centrifugation test to evaluate the assay as an alternative technique for quantifying the intact 146S antigen of both SAT serotypes. In BHK-21 cells, the FMDV 146S antigen of both SAT serotypes was generated. For each serotype, sixteen samples were examined in duplicates using the SDG (10-30%) centrifugation and polyclonal DAS-ELISA at varied time intervals (0, 5, 10, 15 minutes) and dissociation conditions (Intact 146S, Temperature, pH, complete dissociation (CD)). The SDG method identified the intact 146S antigen particles at 254 nm using a Type 11 Optical Unit for a UA-6 absorbance detector, whereas polyclonal DAS-ELISA detected FMDV antigen particles at OD 450 nm using a microplate ELISA reader. The SDG was more specific to the Immunogenically intact 146S antigen, whereas polyclonal DAS-ELISA measured an equivalent reactivity to the intact 146S antigen and the 12S protein components in both SAT serotypes. The polyclonal DAS-ELISA technique was not suitable for quantifying the intact 146S antigen and so could not be used for quantification of the immunogenic 146S antigen component during vaccine production.en_US
dc.description.availabilityUnrestricteden_US
dc.description.degreeMSc (Tropical Animal Health)en_US
dc.description.departmentVeterinary Tropical Diseasesen_US
dc.description.facultyFaculty of Veterinary Scienceen_US
dc.description.sponsorshipARCen_US
dc.description.sponsorshipUPen_US
dc.identifier.citation*en_US
dc.identifier.doi10.25403/UPresearchdata.25133804en_US
dc.identifier.otherA2024en_US
dc.identifier.urihttp://hdl.handle.net/2263/94389
dc.language.isoenen_US
dc.publisherUniversity of Pretoria
dc.rights© 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
dc.subjectUCTDen_US
dc.subjectSucrose Density Gradient (SDG)
dc.subjectPolyclonal DAS-ELISA
dc.subjectComplete dissociation (CD)
dc.subject146S antigen
dc.subject12S protein subunits
dc.subjectFMD virus (FMDV)
dc.titleEvaluation of a DAS-ELISA for quantification of foot-and-mouth disease virus 146S antigen during vaccine preparationen_US
dc.typeMini Dissertationen_US

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