title: [Eukaryotic expression of human IL17-RD-ECD and generation of its monoclonal antibody].
abstract: IL-17 Receptor D (IL-17 RD) is a cytokine receptor that mediates IL-17 signaling and plays an important role in responding to the invasion of extracellular pathogens and many inflammatory and autoimmune diseases such as rheumatoid arthritis. In this study we report the generation of a mouse monoclonal antibody against human IL-17 RD. The recombinant human IL-17RD extracellular domain (hIL-17RD-ECD) was produced in the baculovirus expression system and purified from culture medium of sf9 insect cells. The purified protein was used as a T-dependent antigen to immune Balb/C mice. B cells from the spleen of immunized mice were fused with murine cell SP2/0. Hybridoma cell lines were screened for the production of the monoclonal antibody against hIL-17-RD-ECD using ELISA. A hybridoma cell line 1F8 was found to have a high production of the antibody, which was further confirmed for the specificity by both western blot and ELISA analyses. The monoclonal antibody obtained from hybridoma 1F8 was characterized to be IgG1+Kappa subclass. This study provided a base for the further therapeutic application of the antibody on the autoimmune disease including rheumatoid arthritis.
authors.affiliation: School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Sun
authors.forename: Kunrong
authors.initials: K
authors.lastname: Mei
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Wang
authors.forename: Fangli
authors.initials: F
authors.lastname: Ren
authors.forename: Yongjing
authors.initials: Y
authors.lastname: Xia
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Chang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016367:Baculoviridae; D006801:Humans; D007313:Insecta / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D053722:Receptors, Interleukin-17 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D053722:Receptors, Interleukin-17; D011994:Recombinant Proteins