Generation of a panel of monoclonal antibodies against atypical chemokine receptor CCX-CKR by DNA immunization.

摘要: Abstract Introduction Chemokines are regulated by a family of ‘atypical’ chemokine receptors, D6, DARC and CCX-CKR, each which efficiently internalizes its cognate ligands. Development monoclonal antibodies (MAbs) that would recognize CCX-CKR on the cell surface will be helpful to identify primary CCX-CKR-expressing types analyze fate after ligand binding receptor. Methods We generated IgG MAbs recognizing cell-surface DNA immunization using molecular adjuvant, analyzed epitope recognized MAbs. Then, reactivities with CCX-CKR-transfected cells, also hepatocytes hepatic tumor lines were evaluated. Finally, we tested ligand-like activities MAbs, namely, induction internalization Results A panel reacting expressed was prepared. The small one, consisting only ten but rich in terms diversity Ig isotypes epitopes. Epitope analyses revealed all 10 at least three different, although very close, peptide structures N-terminal domain. Three 2F11, 13E11 14F10, selected represent panel. All applicable for flow cytometry immunoflurescent assays immunoprecipitation. reactivity 2F11 MAb confirmed western blotting. Endogenous expression human demonstrated MAb. Interestingly, B300-19 cells expressing resulted internalization. Discussion This may expected prove valuable further study functions this silent receptor, including those related homeostasis lymphoid growth metastasis cancer.