Preparation of a monoclonal antibody against transporter associated with antigen processing in MHC haplotype SPF ducks
Received:October 30, 2018  
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DOI:10. 3969 / j.issn.1005-4847. 2019. 02. 012
KeyWord:major histocompatibility complex ( MHC) haplotype duck; transporter associated with antigen processing; TAP; monoclonal antibody; specificity
              
AuthorInstitution
王兴童 中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队,兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,哈尔滨
孟兴 中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队,兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,哈尔滨
佟相慧 中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队,兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,哈尔滨
陈洪岩 中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队,兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,哈尔滨
韩凌霞 中国农业科学院哈尔滨兽医研究所实验动物与比较医学团队,兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,哈尔滨
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Abstract:
      Objective A monoclonal antibody (Mab) against transporter associated with antigen processing (TAP)was prepared to study the function of TAP protein in the immunogenetics of ducks. Methods BALB/ c mice were immunizedwith fused Escherichia coli expression products containing peptide-binding region of TAP protein of major histocompatibilitycomplex (MHC) haplotype SPF ducks. Specific monoclonal antibodies were screened by ELISA, and truncated expressionmethod was used to identify the antigenic epitope. An indirect immunofluorescence assay was used to compare the reactivity ofMab against the peripheral blood lymphocytes of ducks and chickens. Immunohistochemistry was used to identify thespecificity of the Mab was detected against SPF chickens, SPF ducks, SPF pigs, common quails and geese. Results A duckTAP-specific monoclonal antibody was obtained and named as “1A6”. The antigenic epitope against 1A6 was identifiedroughly at 297NARHQMLQQAVLDATAGTGMVVQEAI322 by a trimmed expressed fusion protein. Chicken and duck peripheralbloodlymphocytes were tested positive for 1A6 in this indirect immunofluorescence assay. Immunohistochemistry of theduodenum from SPF chickens, SPF ducks, SPF pigs, Corturnix and geese by 1A6 was undertaken. Specific strong signals inimmunohistochemistry assay were detected in the duodenum of chickens and ducks, but not detected in the pigs, quails,geese or negative-control jejunum. Conclusions A monoclonal antibody specific to the TAP protein of chickens and ducks is obtained. It provides a useful material for studies of avian diseases and immunology of poultry.
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