[Preparation of two mouse anti-human PD-1 monoclonal antibodies and identification of their biological characteristics].
OBJECTIVE
To prepare mouse anti-human PD-1 monoclonal antibodies (mAbs) and identify their biological characteristics.


METHODS
The BALB/c mice were immunized with the transfected cell line PD-1/L929. The cells were fused with Sp2/0 using monoclonal antibody techniques and the positive clones were screened by FCS with PD-1/L929. The secreted anti-PD-1 mAbs were identified through rapid isotyping analysis, karyotype analysis, Western blot, competitive inhibition test, indirect immunofluorescence assay, and tumor cell lines detection.


RESULTS
Two mouse anti-human PD-1 hybridomas were obtained and their secreted mAbs were named (1F2 and 5F10). Their biological characteristics suggested that they could recognize a protein with approximate molecular weight 55 000 on PD-1/L929 cell lines and different epitopes. 1F2 could recognize PD-1 molecules expressed on SKHep-1 and 7721 while 5F10 could recognize Raji cells.


CONCLUSIONS
Two mouse anti-human PD-1 hybridoma cell lines and their secreted monoclonal antibodies have been successfully obtained and identified.
Ming
Zhi-jun
ZJ
Institute of Medical Biotechnology, Clinical Immunology Research Laboratory of Jiangsu Province, Suzhou 215123, China. mingzhijun@suda.edu.cn
Wu
Hai-jing
HJ
Zhang
Guang-bo
GB
Hu
Yu-min
YM
Qiu
Yu-hua
YH
Zhang
Xue-guang
XG
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D000918:Antibody Specificity / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000276:immunology*; D002460:Cell Line; D045744:Cell Line, Tumor; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D061026:Programmed Cell Death 1 Receptor
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D051017:Apoptosis Regulatory Proteins; D000939:Epitopes; C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor