Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species
Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and diagnostic tests. Although there have been many reports of such work done for SARS–CoV, these were mainly focused on the immune responses of humans and mice. In this study, we aim...
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| Format: | Article |
| Language: | English |
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Elsevier B.V.
2008
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| Online Access: | http://ir.unimas.my/id/eprint/17687/ http://ir.unimas.my/id/eprint/17687/1/Meng%20Yu.pdf |
| _version_ | 1848838344287453184 |
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| author | Cardosa, Mary Jane |
| author_facet | Cardosa, Mary Jane |
| author_sort | Cardosa, Mary Jane |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and
diagnostic tests. Although there have been many reports of such work done for SARS–CoV, these were mainly focused on the
immune responses of humans and mice. In this study, we aim to search for and compare immunodominant regions of the spike (S)
and nucleocapsid (N) proteins which are recognized by sera from different animal species, including mouse, rat, rabbit, civet, pig
and horse. Twelve overlapping recombinant protein fragments were produced in Escherichia coli, six each for the S and N
proteins, which covered the entire coding region of the two proteins. Using a membrane-strip based Western blot approach, the
reactivity of each antigen fragment against a panel of animal sera was determined. Immunodominant regions containing linear
epitopes, which reacted with sera from all the species tested, were identified for both proteins. The S3 fragment (aa 402–622) and
the N4 fragment (aa 220–336) were the most immunodominant among the six S and N fragments, respectively. Antibodies raised
against the S3 fragment were able to block the binding of a panel of S-specific monoclonal antibodies (mAb) to SARS–CoV in
ELISA, further demonstrating the immunodominance of this region. Based on these findings, one-step competition ELISAs were
established which were able to detect SARS–CoV antibodies from human and at least seven different animal species. Considering
that a large number of animal species are known to be susceptible to SARS–CoV, these assays will be a useful tool to trace the
origin and transmission of SARS–CoV and to minimise the risk of animal-to-human transmission. |
| first_indexed | 2025-11-15T06:54:04Z |
| format | Article |
| id | unimas-17687 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:54:04Z |
| publishDate | 2008 |
| publisher | Elsevier B.V. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-176872022-01-19T08:32:20Z http://ir.unimas.my/id/eprint/17687/ Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species Cardosa, Mary Jane Q Science (General) Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and diagnostic tests. Although there have been many reports of such work done for SARS–CoV, these were mainly focused on the immune responses of humans and mice. In this study, we aim to search for and compare immunodominant regions of the spike (S) and nucleocapsid (N) proteins which are recognized by sera from different animal species, including mouse, rat, rabbit, civet, pig and horse. Twelve overlapping recombinant protein fragments were produced in Escherichia coli, six each for the S and N proteins, which covered the entire coding region of the two proteins. Using a membrane-strip based Western blot approach, the reactivity of each antigen fragment against a panel of animal sera was determined. Immunodominant regions containing linear epitopes, which reacted with sera from all the species tested, were identified for both proteins. The S3 fragment (aa 402–622) and the N4 fragment (aa 220–336) were the most immunodominant among the six S and N fragments, respectively. Antibodies raised against the S3 fragment were able to block the binding of a panel of S-specific monoclonal antibodies (mAb) to SARS–CoV in ELISA, further demonstrating the immunodominance of this region. Based on these findings, one-step competition ELISAs were established which were able to detect SARS–CoV antibodies from human and at least seven different animal species. Considering that a large number of animal species are known to be susceptible to SARS–CoV, these assays will be a useful tool to trace the origin and transmission of SARS–CoV and to minimise the risk of animal-to-human transmission. Elsevier B.V. 2008 Article PeerReviewed text en http://ir.unimas.my/id/eprint/17687/1/Meng%20Yu.pdf Cardosa, Mary Jane (2008) Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species. Journal of Immunological Methods, 331. ISSN 0022-1759 http://www.jimmunol.org/content/jimmunol/173/6/4050.full.pdf doi:10.1016/j.jim.2007.11.009 |
| spellingShingle | Q Science (General) Cardosa, Mary Jane Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title | Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title_full | Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title_fullStr | Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title_full_unstemmed | Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title_short | Determination and application of immunodominant regions of SARS coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| title_sort | determination and application of immunodominant regions of sars coronavirus spike and nucleocapsid proteins recognized by sera from different animal species |
| topic | Q Science (General) |
| url | http://ir.unimas.my/id/eprint/17687/ http://ir.unimas.my/id/eprint/17687/ http://ir.unimas.my/id/eprint/17687/ http://ir.unimas.my/id/eprint/17687/1/Meng%20Yu.pdf |