Title: The preparation and evaluation of the monoclonal antibodies against nucleocapsid protein of human parainfluenza virus type 3
Abstract: Objective To prepare the monoclonal antibodies(MAbs)against nucleocapsid protein(NP)of human parainfluenza virus type 3(HPIV-3)and evaluate the possible use in the development of early diagnosis reagent for HPIV-3 infection.Methods Recombinant nucleocapsid protein from HPIV-3 were induced for expression in BL21 cells and purified with GST affinity agarose column.BALB/C mice were immunized with purified recombinant NP and the antisera were preliminarily tested for sifting the mice with high immune response.The hybridoma cells were constructed from immunized mice spleen cells and SP2/0 cells with PEG1500.The supernatant fluid of hybridoma cells was tested for specific monoclonal antibodies by Enzyme-Linked Immunosorbent Assay(ELISA)and indirect immunofluorescence assay(IFA).Then large yields of the MAbs were induced via cell culture or peritoneal inoculation with hybridoma.The MAbs were evaluated for specific and cross reaction with HPIV-3 and other common respiratory virus in pediatrics,respectively.Results Nine hybridoma cell lines were constructed for excreting specific monoclonal antibodies against recombinant NP.Two of them could recognize natural HPIV-3 at titrations of 1∶1600 and 1∶6400 respectively by IFA.Conclusions The recombinant NP can elicit monoclonal antibodies against natural human parainfluenza virus type 3,and it has the potential to be used in the diagnosis of HPIV-3 infection.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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