[Preparation and identification of monoclonal antibodies against human DcR3].
OBJECTIVE
To prepare monoclonal antibodies(mAbs) against human DcR3 and identify their characterization.


METHODS
BALB/c mice were immunized with purified His-DcR3 protein, and mAbs against DcR3 which prepared by hybridoma technique were purified and identified by their specificity, subtype, titers via ELISA and Western blot.


RESULTS
Five hybridoma cell lines secreting mAbs against human DcR3 were obtained, which were determined as IgG1 subtype and ascites titers of five mAbs against DcR3 reached 1x10(-5)-1x10(-7). Five mAbs were proved to recognize His-DcR3 protein specifically, one of which (1B1) could recognize SW480 cell.


CONCLUSIONS
mAbs against DcR3 with high titers and specificity have been prepared and purified successfully, which laid a foundation for the study of DcR3 expression, distribution in tissu and development of ELISA kit.
Wu
Chun-feng
CF
Department of Respiratory Diseases, Jilin University, Changchun 130041, China. feng1218@sohu.com
Ma
Zhong-sen
ZS
Wang
Xiu-li
XL
Yang
Jun-ling
JL
Yang
Yang
Y
Zhang
Yue
Y
Tumen
Wu-li-ji
WL
Qiao
Jun-hua
JH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D053323:Receptors, Tumor Necrosis Factor, Member 6b / Q000276:immunology*
D000911:Antibodies, Monoclonal; D053323:Receptors, Tumor Necrosis Factor, Member 6b; C505911:TNFRSF6B protein, human