Preparation of PrP-specific Polyclonal Antibody via Immunization of PRNP-knockout Mice with Recombinant Human PrP Protein

作     者：YANG Xue Hua WU Yue Zhang XIAO Kang GAO Li Ping CHEN Dong Dong DONG Xiao Ping SHI Qi YANG Xue Hua;WU Yue Zhang;XIAO Kang;GAO Li Ping;CHEN Dong Dong;DONG Xiao Ping;SHI Qi

作者机构：State Key Laboratory for Infectious Disease Prevention and ControlCollaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases(Zhejiang University)National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Center for Global Public HealthChinese Center for Disease Control and PreventionBeijing 102206China Wuhan Institute of VirologyChinese Academy of ScienceWuhan 430071HubeiChina China Academy of Chinese Medical SciencesBeijing 100700China 

出 版 物：《Biomedical and Environmental Sciences》 (生物医学与环境科学（英文版）)

年 卷 期：2020年第33卷第7期

页      面：493-501页

核心收录：

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学] 

基　　金：National Natural Science Foundation[Grant Nos.81572048 and 81630062] the National Key R&D Program of China[Grant Nos.2018ZX10711001,2016YFC1202700,and 2017YFC1200500] the State Key Laboratory for Infectious Disease Prevention and Control,China CDC[Grant Nos.2019SKLID501,2019SKLID603,and 2019SKLID307] 

主　　题：Prion disease PrP Antibody PRNP-knockout mouse 

摘      要：Objective The definite diagnosis of human and animal prion diseases depends on the examination of special pathological changes and/or detection of PrPSc in the brain tissues of suspected ***,developing methods to obtain PrP antibody with good specificity and sensitivity is fundamental for prion *** We prepared a PrP-specific polyclonal antibody(pAb P54)in a PRNP-knockout mouse model via immunization with recombinant full-length human PrP protein residues 23–***,we verified that pAb in Western blot,immunohistochemistry(IHC),and immunofluorescent(IFA)*** Western blot illustrated that the newly prepared pAb P54 could react with recombinant PrP protein,normal brain PrPC from healthy rodents and humans,and pathological PrPSc in the brains of experimental rodents infected with scrapie and humans infected with different types of prion *** electrophoretic patterns of brain PrPC and PrPSc observed after their reaction with pAb P54 were nearly identical to those produced by commercial PrP monoclonal *** glycosylated PrP molecules in the brain homogenates were clearly demonstrated in the reactions of these molecules with pAb *** assay revealed apparent PrP deposits in the GdnCl-treated brain slices of 139 A-infected mice and 263 K-infected *** tests with pAb P54 also showed clear green signals surrounding blue-stained cell *** The newly prepared pAb P54 demonstrated reliable specificity and sensitivity and,thus,may have potential applications not only in studies of prion biology but also in the diagnosis of human and experimental rodent prion diseases.