Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids
Analysis of the immune response to infection of livestock by foot-and-mouth disease virus (FMDV) is most often reported as the serum antibody response to the virus. While measurement of neutralizing antibody has been sensitive and specific, measurements of the quality of the antibody response are le...
| Autores principales: | , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2019
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/4444 https://www.sciencedirect.com/science/article/pii/S0022175917302144?via%3Dihub https://doi.org/10.1016/j.jim.2017.07.001 |
| _version_ | 1855035324505260032 |
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| author | Kenney, Mary Waters, Ryan A. Rieder, Elizabeth Pega, Juan Franco Perez Filgueira, Daniel Mariano Golde, William T. |
| author_browse | Golde, William T. Kenney, Mary Pega, Juan Franco Perez Filgueira, Daniel Mariano Rieder, Elizabeth Waters, Ryan A. |
| author_facet | Kenney, Mary Waters, Ryan A. Rieder, Elizabeth Pega, Juan Franco Perez Filgueira, Daniel Mariano Golde, William T. |
| author_sort | Kenney, Mary |
| collection | INTA Digital |
| description | Analysis of the immune response to infection of livestock by foot-and-mouth disease virus (FMDV) is most often reported as the serum antibody response to the virus. While measurement of neutralizing antibody has been sensitive and specific, measurements of the quality of the antibody response are less robust. Determining the immunoglobulin (Ig) isotype of the serum antibody response provides a deeper understanding of the biology of the response and more sensitive methods for these assays will facilitate analyses of B cell mediated immunity. We tested the hypothesis that using the virus as the molecular probe could be achieved by adding tags to the surface of the FMDV capsid, and that would enhance sensitivity in assays for anti-FMDV antibody responses. The use of a FLAG-tagged virus in these assays failed to yield improvement whereas chemically biotinylating the virus capsid resulted in significant enhancement of the signal. Here we describe methods using biotinylated virus for measuring anti-viral antibody in serum and antibody secreting cells (ASCs) in blood that are sensitive and specific. Finally, we describe using the biotinylated virus in flow cytometry where such assays should greatly enhance the analysis of anti-virus antibody producing B cells, allowing the investigator to focus on only the FMDV specific B cells when analyzing the development of the B cell response to either infection or vaccination. |
| format | info:ar-repo/semantics/artículo |
| id | INTA4444 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA44442019-02-14T15:55:29Z Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids Kenney, Mary Waters, Ryan A. Rieder, Elizabeth Pega, Juan Franco Perez Filgueira, Daniel Mariano Golde, William T. Virus Fiebre Aftosa Respuesta Inmunológica ELISA Citometría de Flujo (Células) Aphthovirus Immune Response Flow Cytometry Foot and Mouth Disease Virus Biotinylation Biotinilación Analysis of the immune response to infection of livestock by foot-and-mouth disease virus (FMDV) is most often reported as the serum antibody response to the virus. While measurement of neutralizing antibody has been sensitive and specific, measurements of the quality of the antibody response are less robust. Determining the immunoglobulin (Ig) isotype of the serum antibody response provides a deeper understanding of the biology of the response and more sensitive methods for these assays will facilitate analyses of B cell mediated immunity. We tested the hypothesis that using the virus as the molecular probe could be achieved by adding tags to the surface of the FMDV capsid, and that would enhance sensitivity in assays for anti-FMDV antibody responses. The use of a FLAG-tagged virus in these assays failed to yield improvement whereas chemically biotinylating the virus capsid resulted in significant enhancement of the signal. Here we describe methods using biotinylated virus for measuring anti-viral antibody in serum and antibody secreting cells (ASCs) in blood that are sensitive and specific. Finally, we describe using the biotinylated virus in flow cytometry where such assays should greatly enhance the analysis of anti-virus antibody producing B cells, allowing the investigator to focus on only the FMDV specific B cells when analyzing the development of the B cell response to either infection or vaccination. Instituto de Virología Fil: Kenney, Mary. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos Fil: Waters, Ryan A. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos Fil: Rieder, Elizabeth. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos Fil: Pega, Juan Franco. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Perez Filgueira, Daniel Mariano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Golde, William T. United States Department of Agriculture. Agricultural Research Service. Plum Island Animal Disease Center; Estados Unidos 2019-02-14T15:50:37Z 2019-02-14T15:50:37Z 2017-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/4444 https://www.sciencedirect.com/science/article/pii/S0022175917302144?via%3Dihub 0022-1759 https://doi.org/10.1016/j.jim.2017.07.001 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Journal of immunological methods 450 : 1-9. (November 2017) |
| spellingShingle | Virus Fiebre Aftosa Respuesta Inmunológica ELISA Citometría de Flujo (Células) Aphthovirus Immune Response Flow Cytometry Foot and Mouth Disease Virus Biotinylation Biotinilación Kenney, Mary Waters, Ryan A. Rieder, Elizabeth Pega, Juan Franco Perez Filgueira, Daniel Mariano Golde, William T. Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title | Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title_full | Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title_fullStr | Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title_full_unstemmed | Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title_short | Enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot-and-mouth disease virus (FMDV) capsids |
| title_sort | enhanced sensitivity in detection of antiviral antibody responses using biotinylation of foot and mouth disease virus fmdv capsids |
| topic | Virus Fiebre Aftosa Respuesta Inmunológica ELISA Citometría de Flujo (Células) Aphthovirus Immune Response Flow Cytometry Foot and Mouth Disease Virus Biotinylation Biotinilación |
| url | http://hdl.handle.net/20.500.12123/4444 https://www.sciencedirect.com/science/article/pii/S0022175917302144?via%3Dihub https://doi.org/10.1016/j.jim.2017.07.001 |
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