Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies
Filoviruses cause lethal hemorrhagic fever in humans. The filovirus nucleoprotein (NP) is expressed in high abundance in infected cells and is essential for virus replication. To generate anti-filovirus monoclonal antibodies (mAbs) against the NP, mice were immunized with peptides known as B-cell ep...
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doaj-8360f0dbfc0b4067819df6e4e0369f682020-11-25T02:50:44ZengMDPI AGViruses1999-49152019-03-0111325910.3390/v11030259v11030259Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal AntibodiesMd Niaz Rahim0Min Wang1Tong Wang2Shihua He3Bryan D. Griffin4Darwyn Kobasa5Ruifu Yang6Zongmin Du7Xiangguo Qiu8Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, CanadaState Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, ChinaState Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, ChinaSpecial Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, CanadaSpecial Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, CanadaSpecial Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, CanadaState Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, ChinaState Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, ChinaSpecial Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, MB, R3E 3R2, CanadaFiloviruses cause lethal hemorrhagic fever in humans. The filovirus nucleoprotein (NP) is expressed in high abundance in infected cells and is essential for virus replication. To generate anti-filovirus monoclonal antibodies (mAbs) against the NP, mice were immunized with peptides known as B-cell epitopes corresponding to different filovirus NPs, and hybridomas were screened using FLAG-tagged filovirus NP constructs. Numerous mAbs were identified, isotyped, and characterized. The anti-NP mAbs demonstrated different ranges of binding affinities to various filovirus NPs. Most of the clones specifically detected both recombinant and wild-type NPs from different filoviruses, including Ebola (EBOV), Sudan (SUDV), Bundibugyo (BDBV), Marburg (MARV), Tai Forest (TAFV), and Reston (RESTV) viruses in western blot analysis. The mAbs were also able to detect native NPs within the cytoplasm of infected cells by immunofluorescence confocal microscopy. Thus, this panel of mAbs represents an important set of tools that may be potentially useful for diagnosing filovirus infection, characterizing virus replication, and detecting NP–host protein interactions.http://www.mdpi.com/1999-4915/11/3/259mAb (monoclonal antibody)Ebola (EBOV)Sudan (SUDV)Bundibugyo (BDBV)Marburg (MARV)Tai Forest (TAFV)Reston (RESTV) viruses |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Md Niaz Rahim Min Wang Tong Wang Shihua He Bryan D. Griffin Darwyn Kobasa Ruifu Yang Zongmin Du Xiangguo Qiu |
spellingShingle |
Md Niaz Rahim Min Wang Tong Wang Shihua He Bryan D. Griffin Darwyn Kobasa Ruifu Yang Zongmin Du Xiangguo Qiu Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies Viruses mAb (monoclonal antibody) Ebola (EBOV) Sudan (SUDV) Bundibugyo (BDBV) Marburg (MARV) Tai Forest (TAFV) Reston (RESTV) viruses |
author_facet |
Md Niaz Rahim Min Wang Tong Wang Shihua He Bryan D. Griffin Darwyn Kobasa Ruifu Yang Zongmin Du Xiangguo Qiu |
author_sort |
Md Niaz Rahim |
title |
Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies |
title_short |
Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies |
title_full |
Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies |
title_fullStr |
Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies |
title_full_unstemmed |
Generation and Characterization of Anti-Filovirus Nucleoprotein Monoclonal Antibodies |
title_sort |
generation and characterization of anti-filovirus nucleoprotein monoclonal antibodies |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2019-03-01 |
description |
Filoviruses cause lethal hemorrhagic fever in humans. The filovirus nucleoprotein (NP) is expressed in high abundance in infected cells and is essential for virus replication. To generate anti-filovirus monoclonal antibodies (mAbs) against the NP, mice were immunized with peptides known as B-cell epitopes corresponding to different filovirus NPs, and hybridomas were screened using FLAG-tagged filovirus NP constructs. Numerous mAbs were identified, isotyped, and characterized. The anti-NP mAbs demonstrated different ranges of binding affinities to various filovirus NPs. Most of the clones specifically detected both recombinant and wild-type NPs from different filoviruses, including Ebola (EBOV), Sudan (SUDV), Bundibugyo (BDBV), Marburg (MARV), Tai Forest (TAFV), and Reston (RESTV) viruses in western blot analysis. The mAbs were also able to detect native NPs within the cytoplasm of infected cells by immunofluorescence confocal microscopy. Thus, this panel of mAbs represents an important set of tools that may be potentially useful for diagnosing filovirus infection, characterizing virus replication, and detecting NP–host protein interactions. |
topic |
mAb (monoclonal antibody) Ebola (EBOV) Sudan (SUDV) Bundibugyo (BDBV) Marburg (MARV) Tai Forest (TAFV) Reston (RESTV) viruses |
url |
http://www.mdpi.com/1999-4915/11/3/259 |
work_keys_str_mv |
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