SPOT SYNTHESIS OF OVERLAPPING PEPTIDES ON PAPER MEMBRANE SUPPORTS ENABLES THE IDENTIFICATION OF LINEAR MONOCLONAL-ANTIBODY BINDING DETERMINANTS ON MORBILLIVIRUS PHOSPHOPROTEINS

In order to map antigenic domains on the P-protein of morbillivirus, a series of overlapping peptides, representing the P-protein sequences of phocid distemper virus strain 2558/Han88 and canine distemper virus strain Onderstepoort, were synthesized on a paper support by the spot -technique. The reactivity of six monoclonal antibodies with the pepti des was tested in an enzyme immunoassay and compared to their reactivi ty in Western blots and in an ELISA using detergent extracts from viru s-infected cells, Three linear determinants could be localized on the P-protein. Two antibody-binding sites were delineated within the C-ter minal (between amino acids 307-322 and 382-400, respectively), and a t hird one was located on the N-terminal part (amino acids 13-31) of the protein. Fine mapping of this binding site revealed that this was a p art of an antigenic domain. In Western blots, the monoclonal antibodie s reacting with this domain also reacted with a second protein which w as possibly the V-protein.