[Preparation and characterization of monoclonal antibody against DR5].
OBJECTIVE
To prepare monoclonal antibodies(mAb) against DR5 and characterize their properties.


METHODS
BALB/c mice were immunized with DR5, and then mAb was prepared by hybridoma technique. Ig subclass and specificity of mAbs was analyzed by ELISA and flow cytometry, respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA.


RESULTS
Four hybridoma cell lines secreting anti DR5 mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 4 mAbs in ascitic fluid were 1x10(-4) - 5x10(-6). Affinity constant of mAbs were 1x10(9). They recognized 2 different epitopes on DR5 molecule.


CONCLUSIONS
Four mAbs against DR5 are prepared successfully, which provides useful reagent for clinical diagnosis and further research.
Bai
Hui-ling
HL
Research Institute of Immunology, Henan University, Kaifeng 475001, China. kfbaihling369@tom.com
Zhao
Yue-ping
YP
Liu
Guang-chao
GC
Ma
Yuan-fang
YF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D019169:Jurkat Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000276:immunology*; D012995:Solubility
D000911:Antibodies, Monoclonal; D000939:Epitopes; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand