PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO HUMAN CILIARY NEUROTROPHIC FACTOR WITH DEFINED EPITOPE RECOGNITION

Ten mouse hybridoma lines producing monoclonal antibodies (Mabs) again st recombinant human ciliary neurotrophic factor (rhCNTF) have been ob tained. Two monoclonal antibodies belonging to the IgG1 class were sel ected and characterized. Their specificity was established by ELISA an d Western blotting. Epitopes recognized by the two Mabs were investiga ted with ELISA and Western blotting by using rhCNTF mutants, rhCNTF fr agments and synthetic peptides mimicking different portions of the CNT F molecule. The carboxy-terminal part of the CNTF and particularly the sequence between aa 150 and 159 appeared to constitute the immunodomi nant group. The fact that certain amino acid sequences of CNTF are con served among species was utilized to examine the crossreactivity patte rns of the two Mabs with rat sciatic nerve CNTF by Western blotting an d immunohistochemistry. These antibodies will be useful for studying t he distribution of CNTF in the nervous system and in developing an enz yme-linked immunosorbent assay for the quantitative determinations of CNTF in various neuropathologies.