Development of monoclonal anti-aryl hydrocarbon receptor (AHR) antibodies and biopanning for peptide ligands to the AHR
Alles, Neva J
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The aryl hydrocarbon receptor (AHR) is a highly conserved DNA transcription factor liganded by exogenous synthetic ligands. The prototypical ligand is 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin), a halogenated aromatic hydrocarbon (HAH). Toxic levels of HAH exposure may cause immunosuppression, tumor promotion, endocrine disruption, developmental and reproductive toxicity. Endogenous AHR ligands, with related biological roles, are not known but are suspected. This research attempted to identify possible endogenous ligands utilizing a high through-put biopanning technique, with bacteriophage (phage) peptide libraries. Isolated peptides may be related to the endogenous ligand(s). The development of monoclonal anti-AHR antibodies was also undertaken to enable the capture of a higher concentration of AHR and aid in better receptor orientation for biopanning. Monoclonal AHR antibodies were produced to an amino (N) terminus region and a carboxy (C) terminus region of the AHR protein through immunization of Balb/C, C57BL/6 and AHR normal (+/+) and knockout (-/-) Bradfield strain mice.