![]() The major impediment in generating functional Abs for Ab drug development may be the difficulty in obtaining native forms of mAgs to immunize animals and follow the Ab characterization process. Up to 50–60% of the current drugs on the market are GPCR-targeted, but only two are Ab drugs 3. Membrane antigens (mAgs), such as G protein-coupled receptors (GPCRs) that constitute a large family of protein receptors are attractive and important targets for antibody (Ab) therapy in various diseases 1, 2. Therefore, the cell adjuvant could preserve the native conformation of mAgs and exhibit anti-mAg Ab stimulatory ability, providing a more convenient and effective method to generate functional Abs, thus possibly accelerating Ab drug development. Several hybridoma clones also exhibited selective binding to 293 A/CXCR2 cells. The immune serum exhibited relatively higher binding to xenogeneic 293 A/CXCR2 cells than 293 A cells (~3.5-fold). A G-protein coupled receptor (GPCR), CXCR2, was then transiently expressed on 3T3/mGM-CSF cell adjuvant to immunize mice. We found that 3T3/mGM-CSF cells stimulated higher specific anti-EVI2B Ab response in the immunized mice than the other cell adjuvants. We then transiently expressed ecotropic viral integration site 2B (EVI2B) on the adjuvants and used them to directly immunize BALB/c mice. We established cell adjuvants by expressing membrane-bound cytokines (mIL-2, mIL-18, or mGM-CSF) on BALB/3T3 cells, which were effective in stimulating splenocyte proliferation in vitro. Then, mAgs-expressing xenogeneic cells were used for Ab characterization to reduce non-specific binding. The membrane-bound cytokines were used as immune stimulators to enhance specific Ab responses against the desired mAgs. In this study, we present a platform in which human mAgs were expressed in native form on cell adjuvants made with membrane-bound cytokines that were then used immunize syngeneic mice directly. However, native mAgs are not easily prepared, causing difficulties in acquiring functional Abs. Membrane antigens (mAgs) are important targets for the development of antibody (Ab) drugs. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |