Cargando…

Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)

The domestic ferret (Mustela putorius furo) serves as an animal model for the study of several viruses that cause human disease, most notably influenza. Despite the importance of this animal model, characterization of the immune response by flow cytometry (FCM) is severely hampered due to the limite...

Descripción completa

Detalles Bibliográficos
Autores principales: Kirchenbaum, Greg A., Ross, Ted M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5329684/
https://www.ncbi.nlm.nih.gov/pubmed/28286781
http://dx.doi.org/10.1155/2017/5874572
_version_ 1782511103494848512
author Kirchenbaum, Greg A.
Ross, Ted M.
author_facet Kirchenbaum, Greg A.
Ross, Ted M.
author_sort Kirchenbaum, Greg A.
collection PubMed
description The domestic ferret (Mustela putorius furo) serves as an animal model for the study of several viruses that cause human disease, most notably influenza. Despite the importance of this animal model, characterization of the immune response by flow cytometry (FCM) is severely hampered due to the limited number of commercially available reagents. To begin to address this unmet need and to facilitate more in-depth study of ferret B cells including the identification of antibody-secreting cells, eight unique murine monoclonal antibodies (mAb) with specificity for ferret immunoglobulin (Ig) were generated using conventional B cell hybridoma technology. These mAb were screened for reactivity against ferret peripheral blood mononuclear cells by FCM and demonstrate specificity for CD79β(+) B cells. Several of these mAb are specific for the light chain of surface B cell receptor (BCR) and enable segregation of kappa and lambda B cells. Additionally, a mAb that yielded surface staining of nearly all surface BCR positive cells (i.e., pan ferret Ig) was generated. Collectively, these MαF-Ig mAb offer advancement compared to the existing portfolio of polyclonal anti-ferret Ig detection reagents and should be applicable to a wide array of immunologic assays including the identification of antibody-secreting cells by FCM.
format Online
Article
Text
id pubmed-5329684
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Hindawi Publishing Corporation
record_format MEDLINE/PubMed
spelling pubmed-53296842017-03-12 Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo) Kirchenbaum, Greg A. Ross, Ted M. J Immunol Res Research Article The domestic ferret (Mustela putorius furo) serves as an animal model for the study of several viruses that cause human disease, most notably influenza. Despite the importance of this animal model, characterization of the immune response by flow cytometry (FCM) is severely hampered due to the limited number of commercially available reagents. To begin to address this unmet need and to facilitate more in-depth study of ferret B cells including the identification of antibody-secreting cells, eight unique murine monoclonal antibodies (mAb) with specificity for ferret immunoglobulin (Ig) were generated using conventional B cell hybridoma technology. These mAb were screened for reactivity against ferret peripheral blood mononuclear cells by FCM and demonstrate specificity for CD79β(+) B cells. Several of these mAb are specific for the light chain of surface B cell receptor (BCR) and enable segregation of kappa and lambda B cells. Additionally, a mAb that yielded surface staining of nearly all surface BCR positive cells (i.e., pan ferret Ig) was generated. Collectively, these MαF-Ig mAb offer advancement compared to the existing portfolio of polyclonal anti-ferret Ig detection reagents and should be applicable to a wide array of immunologic assays including the identification of antibody-secreting cells by FCM. Hindawi Publishing Corporation 2017 2017-02-14 /pmc/articles/PMC5329684/ /pubmed/28286781 http://dx.doi.org/10.1155/2017/5874572 Text en Copyright © 2017 Greg A. Kirchenbaum and Ted M. Ross. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Kirchenbaum, Greg A.
Ross, Ted M.
Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title_full Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title_fullStr Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title_full_unstemmed Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title_short Generation of Monoclonal Antibodies against Immunoglobulin Proteins of the Domestic Ferret (Mustela putorius furo)
title_sort generation of monoclonal antibodies against immunoglobulin proteins of the domestic ferret (mustela putorius furo)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5329684/
https://www.ncbi.nlm.nih.gov/pubmed/28286781
http://dx.doi.org/10.1155/2017/5874572
work_keys_str_mv AT kirchenbaumgrega generationofmonoclonalantibodiesagainstimmunoglobulinproteinsofthedomesticferretmustelaputoriusfuro
AT rosstedm generationofmonoclonalantibodiesagainstimmunoglobulinproteinsofthedomesticferretmustelaputoriusfuro