title: [Preparation and characterization of three novel monoclonal antibodies against human PD-L1].
abstract: OBJECTIVE
  To prepare a functional mouse anti-human PD-L1 monoclonal antibody and to characterize its biological activities.
  
  
  METHODS
  A stable human PD-L1 transfected cell line L929/PD-L1 was used as an antigen to immunize BALB/c mice. By means of the cell fusion technique, multiple cell subcloning, repeated screening with L929/ PD-L1 as target cells and the L929/mock cells used as the negative control, the hybridomas specifically secreting mouse anti-PD-L1 monoclonal antibodies were generated. Then its biological characterization was investigated by rapid murine Ig-subclass typing, Western blotting, indirect immune of luorescene assay, mutual competitive inhibition test. By means of MTT incorporation assay, detected the infection of mAb to T cell proliferation. Three mouse anti-human PD-L1 monoclonal antibodies were generated, named as 11G8, 2G5 and 5C3.
  
  
  RESULTS
  The results of characterization study showed that the monoclonal antibodies could recognize the PD-L1 on the activated T cells. The mAbs could promote T cells proliferation.
  
  
  CONCLUSIONS
  It is evident that the functional monoclonal antibodies for human PD-L1 have been generated, and it would provide the initial material for further study on the role of PD-1/PD-L1 signaling pathways.
authors.affiliation: Institute of Medical Biotechnology Soochow University, Suzhou, China. su-zhouying@163.com
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.forename: Yong-jing
authors.initials: YJ
authors.lastname: Chen
authors.forename: Li-xiong
authors.initials: LX
authors.lastname: Bai
authors.forename: Geng-chao
authors.initials: GC
authors.lastname: Zhu
authors.forename: Xue-feng
authors.initials: XF
authors.lastname: Wang
authors.forename: Xue-guang
authors.initials: XG
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D060890:B7-H1 Antigen / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D002459:Cell Fusion / Q000379:methods; D045744:Cell Line, Tumor; D000939:Epitopes / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D060890:B7-H1 Antigen; C423236:CD274 protein, human; D000939:Epitopes