Eukaryotic expression of human IL17-RD-ECD and generation of its monoclonal antibody.
- Author:
Xiaojun SUN
1
;
Kunrong MEI
;
Yinyin WANG
;
Fangli REN
;
Yongjing XIA
;
Zhijie CHANG
Author Information
1. School of Medicine, Tsinghua University, Beijing 100084, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
biosynthesis;
Baculoviridae;
Humans;
Insecta;
genetics;
metabolism;
Mice;
Mice, Inbred BALB C;
Receptors, Interleukin-17;
biosynthesis;
genetics;
immunology;
Recombinant Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2011;27(10):1507-1512
- CountryChina
- Language:Chinese
-
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.
