[The preparation and identification on monoclonal antibody to platelet-derived growth factor receptor].
OBJECTIVE
Through genetic engineering to produce the fusion protein of glutathione S-transferase (GST) linked to amino-terminal end of platelet-derived growth factor receptor (PDGFR), and to prepare the bioactive monoclonal antibody.


METHODS
With taking GST-PDGFR-N fusion protein as immunogen, the anti-PDGFR monoclonal antibody was produced by using the hybridoma technique, of which then the antigen binding characteristic was identified by indirect enzyme-linked immunosorbent assay (ELISA), Western blot and immunohistochemistry methods.


RESULTS
Two cell strains of hybridoma were obtained and named as 3B12F5 and 3C6H7C11 which secreted the anti-PDGFR monoclonal antibody, of which the class and subtype identification demonstrated both strains to produce all type of IgG1. The indirect ELISA result showed that the titers of ascites fluid which two hybridoma induced were 1 : 102400 and 1 : 25600. Western blot demonstrated that the two antibodies could recognize specifically the immunogen on PDGFR and U251 cell line. The cell immunohistochemistry proved that the antibody could recognize the expressed PDGFR antigens of neurogliocytoma U251 and bladder carcinoma BIU87 cell lines.


CONCLUSIONS
We prepare successfully the PDGFR monoclonal antibody and provide a useful tool for researching on the PDGFR expression and clinical detection.
Wang
Zhen-da
ZD
Department of Immunology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Fang
Li
L
Wei
Da-peng
DP
Zhang
Chong-jie
CJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D000918:Antibody Specificity; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D006825:Hybridomas; D007074:Immunoglobulin G / Q000032:analysis / Q000235:genetics / Q000276:immunology; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D017479:Receptors, Platelet-Derived Growth Factor / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D017479:Receptors, Platelet-Derived Growth Factor