title: [Preparation and identification of a monoclonal antibody against human PD-L1].
abstract: OBJECTIVE
  To prepare mouse anti-human PD-L1 monoclonal antibodies (mAbs) and identify their biological characteristics.
  
  
  METHODS
  BALB/c mice were immunized with recombinant GST-PDL1 protein,and the strain of hybridoma cell secreting anti-PD-L1 mAb was obtained. The specificity of anti-PD-L1 mAb was checked and evaluated with ELISA and Western blot. Its titer, immunoglobulin subtype were also measured. The combining capacity of anti-PD-L1 mAb with PD-L1 on MDA-MB-231 cells, which had been induced with IFN-gamma for 72 hours, was identified through immunohistochemistry and flow cytometry. The peripheral blood mononuclear cell was labeled with CFSE, and the effect of anti-PD-L1 mAb on the activation and proliferation of the lymphocyte induced with antigens was evaluated by flow cytometry analysis.
  
  
  RESULTS
  A strain of hybridoma cell secreting anti-PD-L1 mAb was successfully obtained and identified belong to IgG1 subtype. Its titers in cultural supernatant and ascetic fluid were 1:6400 and 1:102400, respectively, by ELISA. The purified mAbs showed good affinity and specificity against GST-PDL1 protein in Western blot, also could block the PD-1/PD-L1 pathway and promote the proliferation of lymphocyte induced with antigens.
  
  
  CONCLUSIONS
  Hyperactivity mouse anti-human PD-L1 hybridoma cell lines and their secreted monoclonal antibodies have been successfully obtained and identified.
authors.affiliation: State Key Laboratory of Oral Disease, Sichuan University, Chengdu 610041, China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Chen
authors.forename: Li-na
authors.initials: LN
authors.lastname: Dai
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Feng
authors.forename: Xiao-yu
authors.initials: XY
authors.lastname: Li
authors.forename: Si-xiu
authors.initials: SX
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D060890:B7-H1 Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D060890:B7-H1 Antigen; C423236:CD274 protein, human; D011993:Recombinant Fusion Proteins