TY - JOUR
T1 - Characterization of monoclonal antibodies against hantavirus nucleocapsid protein and their use for immunohistochemistry on rodent and human samples
AU - Kucinskaite-Kodze, Indre
AU - Petraityte-Burneikiene, Rasa
AU - Zvirbliene, Aurelija
AU - Hjelle, Brian
AU - Medina, Rafael A.
AU - Gedvilaite, Alma
AU - Razanskiene, Ausra
AU - Schmidt-Chanasit, Jonas
AU - Mertens, Marc
AU - Padula, Paula
AU - Sasnauskas, Kestutis
AU - Ulrich, Rainer G.
N1 - Funding Information:
The authors thank Detlev H. Krüger and Helga Meisel (Berlin, Germany) for support at the initial stage of work, Connie S. Schmaljohn (Fort Detrick, MD, USA) for provision of the PUUV-P360 N protein-encoding plasmid, Gereon Schares (Wusterhausen, Germany) for provision of negative control mAbs raised against Neospora caninum surface proteins and Lieselotte Minke (Wusterhausen) for excellent technical assistance. The experiments were supported by grants from DLR-Internationales Büro des BMBF (grant LTU 02/001), from the Lithuanian State and Study Foundation (project B-16/2006) and United States Public Health Services grant U19 AI45452.
PY - 2011/2
Y1 - 2011/2
N2 - Monoclonal antibodies are important tools for various applications in hantavirus diagnostics. Recently, we generated Puumala virus (PUUV)-reactive monoclonal antibodies (mAbs) by immunisation of mice with chimeric polyomavirus-derived virus-like particles (VLPs) harbouring the 120-amino-acid-long amino-terminal region of the PUUV nucleocapsid (N) protein. Here, we describe the generation of two mAbs by co-immunisation of mice with hexahistidine-tagged full-length N proteins of Sin Nombre virus (SNV) and Andes virus (ANDV), their characterization by different immunoassays and comparison with the previously generated mAbs raised against a segment of PUUV N protein inserted into VLPs. All of the mAbs reacted strongly in ELISA and western blot tests with the antigens used for immunization and cross-reacted to varying extents with N proteins of other hantaviruses. All mAbs raised against a segment of the PUUV N protein presented on chimeric VLPs and both mAbs raised against the full length AND/SNV N protein reacted with Vero cells infected with different hantaviruses. The reactivity of mAbs with native viral nucleocapsids was also confirmed by their reactivity in immunohistochemistry assays with kidney tissue specimens from experimentally SNV-infected rodents and human heart tissue specimens from hantavirus cardiopulmonary syndrome patients. Therefore, the described mAbs represent useful tools for the immunodetection of hantavirus infection.
AB - Monoclonal antibodies are important tools for various applications in hantavirus diagnostics. Recently, we generated Puumala virus (PUUV)-reactive monoclonal antibodies (mAbs) by immunisation of mice with chimeric polyomavirus-derived virus-like particles (VLPs) harbouring the 120-amino-acid-long amino-terminal region of the PUUV nucleocapsid (N) protein. Here, we describe the generation of two mAbs by co-immunisation of mice with hexahistidine-tagged full-length N proteins of Sin Nombre virus (SNV) and Andes virus (ANDV), their characterization by different immunoassays and comparison with the previously generated mAbs raised against a segment of PUUV N protein inserted into VLPs. All of the mAbs reacted strongly in ELISA and western blot tests with the antigens used for immunization and cross-reacted to varying extents with N proteins of other hantaviruses. All mAbs raised against a segment of the PUUV N protein presented on chimeric VLPs and both mAbs raised against the full length AND/SNV N protein reacted with Vero cells infected with different hantaviruses. The reactivity of mAbs with native viral nucleocapsids was also confirmed by their reactivity in immunohistochemistry assays with kidney tissue specimens from experimentally SNV-infected rodents and human heart tissue specimens from hantavirus cardiopulmonary syndrome patients. Therefore, the described mAbs represent useful tools for the immunodetection of hantavirus infection.
UR - https://www.scopus.com/pages/publications/79952243353
U2 - 10.1007/s00705-010-0879-6
DO - 10.1007/s00705-010-0879-6
M3 - Article
C2 - 21161552
AN - SCOPUS:79952243353
SN - 0304-8608
VL - 156
SP - 443
EP - 456
JO - Archives of Virology
JF - Archives of Virology
IS - 3
ER -