{"id":{"repo_id":"oxford-brookes","oai_identifier":"tle:59211a2b-bbad-4590-a469-b0e0b60d7d06:d6bd9758-527a-46cd-bfe2-c433766e8fca:1"},"canonical_url":"https://search.dev.ndltd.org/etd/oxford-brookes/tle:59211a2b-bbad-4590-a469-b0e0b60d7d06:d6bd9758-527a-46cd-bfe2-c433766e8fca:1","repository":{"repo_id":"oxford-brookes","name":"Oxford Brookes University","base_url":"https://radar.brookes.ac.uk/radar/oai"},"display":{"title":"Baculovirus surface display of influenza virus haemagglutinin and its potential as a vaccine","abstract":"Based on data obtained from the World Health Organization (WHO), influenza (or flu) is responsible for highly contagious, acute respiratory diseases in humans. These diseases are caused by influenza viruses and result in 250,000 to 500,000 deaths globally every year from seasonal flu. In addition, outbreaks of new influenza subtypes occur that on occasion quickly spread to become an epidemic or pandemic. Prevention and control of the spread of these diseases are mainly based on an effective vaccination regime. To date, the vast majority of influenza vaccines are manufactured using conventional technology based on embryonated hen’s eggs that takes several months to produce. However, the threat of influenza virus outbreaks emphasizes the need for a novel, rapid vaccine technology that can respond quickly to stop or reduce the threat of epidemic and pandemic influenza. Sub-unit vaccines based on baculovirus surface display technology are an option for the development and generation of safe human vaccines due to the lack of pre-existing immunity, toxicity and ability to replicate in mammalian systems. Furthermore, this technology may provide an efficient way to produce a large amount of influenza vaccine within a short time frame with a relatively low cost. Haemagglutinin (HA), the surface glycoprotein of influenza virus, is the preferable target for recombinant vaccine studies as it is known to be responsible for new influenza subtypes that may cause pandemic outbreaks and is the key immunogenic protein that can elicit protective neutralising antibody in the host. The research in this thesis was focused on investigating baculovirus surface display technology as a novel approach to develop a recombinant influenza vaccine. Several surface display expression vectors were constructed, using either a truncated or full-length HA, to optimise glycoprotein incorporation into the budded virus (BV) envelope of three recombinant baculovirus variants: BacPAK6, BacPAK6 (high-titre) and flashBACULTRA (FBU). Surface display of HA was found to be improved when the native HA signal peptide and transmembrane domain (TMD) were used; replacement of these domains with baculovirus GP64 signal peptide and TMD reduced HA display. The displayed HA was shown to be biologically active by haemadsorption assay and haemagglutination of chicken red blood cells (RBCs). Analysis of HA display by immunoblotting suggested that HA was incorporated into the BV surface envelope in recombinant virus-infected Sf9 and Tni Hi5 cells and that the levels of display varied between the expression vectors and virus promoters tested. This thesis demonstrates the benefits of using the FBU vector with the polyhedrin gene promoter in Tni Hi5 cells for producing HA incorporated into BV. Interestingly, use of the late p6.9 promoter did not result in increased levels of HA displayed in BV. Initial investigations also demonstrated the promise of RNAi technology to improve HA display by reducing GP64 synthesis and incorporation into the BV surface. The ability of baculovirus displayed HA to elicit an antibody response was assessed in an initial immunisation study using BALB/c mice. Preliminary results showed a strong immune response using ELISA and the antibodies generated were able to prevent haemagglutination in an inhibition assay. Overall, this study suggests that baculovirus display of HA is efficacious and may offer a novel approach for rapid and large-scale production of influenza subunit vaccines to control influenza outbreaks.","abstract_html":"Based on data obtained from the World Health Organization (WHO), influenza (or flu) is responsible for highly contagious, acute respiratory diseases in humans. These diseases are caused by influenza viruses and result in 250,000 to 500,000 deaths globally every year from seasonal flu. In addition, outbreaks of new influenza subtypes occur that on occasion quickly spread to become an epidemic or pandemic. Prevention and control of the spread of these diseases are mainly based on an effective vaccination regime. To date, the vast majority of influenza vaccines are manufactured using conventional technology based on embryonated hen’s eggs that takes several months to produce. However, the threat of influenza virus outbreaks emphasizes the need for a novel, rapid vaccine technology that can respond quickly to stop or reduce the threat of epidemic and pandemic influenza. Sub-unit vaccines based on baculovirus surface display technology are an option for the development and generation of safe human vaccines due to the lack of pre-existing immunity, toxicity and ability to replicate in mammalian systems. Furthermore, this technology may provide an efficient way to produce a large amount of influenza vaccine within a short time frame with a relatively low cost. Haemagglutinin (HA), the surface glycoprotein of influenza virus, is the preferable target for recombinant vaccine studies as it is known to be responsible for new influenza subtypes that may cause pandemic outbreaks and is the key immunogenic protein that can elicit protective neutralising antibody in the host. The research in this thesis was focused on investigating baculovirus surface display technology as a novel approach to develop a recombinant influenza vaccine. Several surface display expression vectors were constructed, using either a truncated or full-length HA, to optimise glycoprotein incorporation into the budded virus (BV) envelope of three recombinant baculovirus variants: BacPAK6, BacPAK6 (high-titre) and flashBACULTRA (FBU). Surface display of HA was found to be improved when the native HA signal peptide and transmembrane domain (TMD) were used; replacement of these domains with baculovirus GP64 signal peptide and TMD reduced HA display. The displayed HA was shown to be biologically active by haemadsorption assay and haemagglutination of chicken red blood cells (RBCs). Analysis of HA display by immunoblotting suggested that HA was incorporated into the BV surface envelope in recombinant virus-infected Sf9 and Tni Hi5 cells and that the levels of display varied between the expression vectors and virus promoters tested. This thesis demonstrates the benefits of using the FBU vector with the polyhedrin gene promoter in Tni Hi5 cells for producing HA incorporated into BV. Interestingly, use of the late p6.9 promoter did not result in increased levels of HA displayed in BV. Initial investigations also demonstrated the promise of RNAi technology to improve HA display by reducing GP64 synthesis and incorporation into the BV surface. The ability of baculovirus displayed HA to elicit an antibody response was assessed in an initial immunisation study using BALB/c mice. Preliminary results showed a strong immune response using ELISA and the antibodies generated were able to prevent haemagglutination in an inhibition assay. Overall, this study suggests that baculovirus display of HA is efficacious and may offer a novel approach for rapid and large-scale production of influenza subunit vaccines to control influenza outbreaks.","abstract_has_math":false,"creators":["Alakeely, Riyadh Abduljabbar Abdulsahib"],"institution":"Oxford Brookes University","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["King, Linda","Possee, Robert","Irons, Sarah"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2018,"date_issued":"2018","date_published":"2018","updated_at":"2026-07-24T03:43:24Z","subjects":[],"languages":["en"],"rights":["All rights reserved"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://doi.org/10.24384/S7N5-8P97","outbound_label":"DOI","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Alakeely, Riyadh Abduljabbar Abdulsahib","King, Linda","Possee, Robert","Irons, Sarah"]},{"key":"dc:creator","label":"Author","values":["Alakeely, Riyadh Abduljabbar Abdulsahib"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2018"]},{"key":"dc:publisher","label":"Institution","values":["Oxford Brookes University"]},{"key":"dc:type","label":"Dc Type","values":["thesis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["All rights reserved"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://doi.org/10.24384/S7N5-8P97","https://radar.brookes.ac.uk/radar/file/59211a2b-bbad-4590-a469-b0e0b60d7d06/1/fulltext.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Based on data obtained from the World Health Organization (WHO), influenza (or flu) is responsible for highly contagious, acute respiratory diseases in humans. These diseases are caused by influenza viruses and result in 250,000 to 500,000 deaths globally every year from seasonal flu. In addition, outbreaks of new influenza subtypes occur that on occasion quickly spread to become an epidemic or pandemic. Prevention and control of the spread of these diseases are mainly based on an effective vaccination regime. To date, the vast majority of influenza vaccines are manufactured using conventional technology based on embryonated hen’s eggs that takes several months to produce. However, the threat of influenza virus outbreaks emphasizes the need for a novel, rapid vaccine technology that can respond quickly to stop or reduce the threat of epidemic and pandemic influenza. Sub-unit vaccines based on baculovirus surface display technology are an option for the development and generation of safe human vaccines due to the lack of pre-existing immunity, toxicity and ability to replicate in mammalian systems. Furthermore, this technology may provide an efficient way to produce a large amount of influenza vaccine within a short time frame with a relatively low cost. Haemagglutinin (HA), the surface glycoprotein of influenza virus, is the preferable target for recombinant vaccine studies as it is known to be responsible for new influenza subtypes that may cause pandemic outbreaks and is the key immunogenic protein that can elicit protective neutralising antibody in the host. The research in this thesis was focused on investigating baculovirus surface display technology as a novel approach to develop a recombinant influenza vaccine. Several surface display expression vectors were constructed, using either a truncated or full-length HA, to optimise glycoprotein incorporation into the budded virus (BV) envelope of three recombinant baculovirus variants: BacPAK6, BacPAK6 (high-titre) and flashBACULTRA (FBU). Surface display of HA was found to be improved when the native HA signal peptide and transmembrane domain (TMD) were used; replacement of these domains with baculovirus GP64 signal peptide and TMD reduced HA display. The displayed HA was shown to be biologically active by haemadsorption assay and haemagglutination of chicken red blood cells (RBCs). Analysis of HA display by immunoblotting suggested that HA was incorporated into the BV surface envelope in recombinant virus-infected Sf9 and Tni Hi5 cells and that the levels of display varied between the expression vectors and virus promoters tested. This thesis demonstrates the benefits of using the FBU vector with the polyhedrin gene promoter in Tni Hi5 cells for producing HA incorporated into BV. Interestingly, use of the late p6.9 promoter did not result in increased levels of HA displayed in BV. Initial investigations also demonstrated the promise of RNAi technology to improve HA display by reducing GP64 synthesis and incorporation into the BV surface. The ability of baculovirus displayed HA to elicit an antibody response was assessed in an initial immunisation study using BALB/c mice. Preliminary results showed a strong immune response using ELISA and the antibodies generated were able to prevent haemagglutination in an inhibition assay. Overall, this study suggests that baculovirus display of HA is efficacious and may offer a novel approach for rapid and large-scale production of influenza subunit vaccines to control influenza outbreaks."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Baculovirus surface display of influenza virus haemagglutinin and its potential as a vaccine"]}]}],"canonical_facts":{"dc:contributor":["Alakeely, Riyadh Abduljabbar Abdulsahib","King, Linda","Possee, Robert","Irons, Sarah"],"dc:creator":["Alakeely, Riyadh Abduljabbar Abdulsahib"],"dc:date":["2018"],"dc:description":["Based on data obtained from the World Health Organization (WHO), influenza (or flu) is responsible for highly contagious, acute respiratory diseases in humans. These diseases are caused by influenza viruses and result in 250,000 to 500,000 deaths globally every year from seasonal flu. In addition, outbreaks of new influenza subtypes occur that on occasion quickly spread to become an epidemic or pandemic. Prevention and control of the spread of these diseases are mainly based on an effective vaccination regime. To date, the vast majority of influenza vaccines are manufactured using conventional technology based on embryonated hen’s eggs that takes several months to produce. However, the threat of influenza virus outbreaks emphasizes the need for a novel, rapid vaccine technology that can respond quickly to stop or reduce the threat of epidemic and pandemic influenza. Sub-unit vaccines based on baculovirus surface display technology are an option for the development and generation of safe human vaccines due to the lack of pre-existing immunity, toxicity and ability to replicate in mammalian systems. Furthermore, this technology may provide an efficient way to produce a large amount of influenza vaccine within a short time frame with a relatively low cost. Haemagglutinin (HA), the surface glycoprotein of influenza virus, is the preferable target for recombinant vaccine studies as it is known to be responsible for new influenza subtypes that may cause pandemic outbreaks and is the key immunogenic protein that can elicit protective neutralising antibody in the host. The research in this thesis was focused on investigating baculovirus surface display technology as a novel approach to develop a recombinant influenza vaccine. Several surface display expression vectors were constructed, using either a truncated or full-length HA, to optimise glycoprotein incorporation into the budded virus (BV) envelope of three recombinant baculovirus variants: BacPAK6, BacPAK6 (high-titre) and flashBACULTRA (FBU). Surface display of HA was found to be improved when the native HA signal peptide and transmembrane domain (TMD) were used; replacement of these domains with baculovirus GP64 signal peptide and TMD reduced HA display. The displayed HA was shown to be biologically active by haemadsorption assay and haemagglutination of chicken red blood cells (RBCs). Analysis of HA display by immunoblotting suggested that HA was incorporated into the BV surface envelope in recombinant virus-infected Sf9 and Tni Hi5 cells and that the levels of display varied between the expression vectors and virus promoters tested. This thesis demonstrates the benefits of using the FBU vector with the polyhedrin gene promoter in Tni Hi5 cells for producing HA incorporated into BV. Interestingly, use of the late p6.9 promoter did not result in increased levels of HA displayed in BV. Initial investigations also demonstrated the promise of RNAi technology to improve HA display by reducing GP64 synthesis and incorporation into the BV surface. The ability of baculovirus displayed HA to elicit an antibody response was assessed in an initial immunisation study using BALB/c mice. Preliminary results showed a strong immune response using ELISA and the antibodies generated were able to prevent haemagglutination in an inhibition assay. Overall, this study suggests that baculovirus display of HA is efficacious and may offer a novel approach for rapid and large-scale production of influenza subunit vaccines to control influenza outbreaks."],"dc:format":["application/pdf"],"dc:identifier":["https://doi.org/10.24384/S7N5-8P97","https://radar.brookes.ac.uk/radar/file/59211a2b-bbad-4590-a469-b0e0b60d7d06/1/fulltext.pdf"],"dc:language":["en"],"dc:publisher":["Oxford Brookes University"],"dc:rights":["All rights reserved"],"dc:title":["Baculovirus surface display of influenza virus haemagglutinin and its potential as a vaccine"],"dc:type":["thesis"]},"updated_at":"2026-07-24T03:43:24Z"}