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 peptides was tested in an enzyme immunoassay and compared to their reactivity in Western blots and in an ELISA using detergent extracts from virus-infected cells. Three linear determinants could be localized on the P-protein. Two antibody-binding sites were delineated within the C-terminal (between amino acids 307-322 and 382-400, respectively), and a third 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 part of an antigenic domain. In Western blots, the monoclonal antibodies reacting with this domain also reacted with a second protein which was possibly the V-protein.
Martens
W
W
Institute of Virology, Hannover Veterinary School, Germany.
Greiser-Wilke
I
I
Harder
T C
TC
Dittmar
K
K
Frank
R
R
Orvell
C
C
Moennig
V
V
Liess
B
B
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000918:Antibody Specificity; D002522:Chlorocebus aethiops; D004216:Distemper / Q000821:virology*; D004217:Distemper Virus, Canine / Q000378:metabolism*; D018104:Distemper Virus, Phocine / Q000378:metabolism*; D000939:Epitopes / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018185:Morbillivirus Infections / Q000821:virology*; D010750:Phosphoproteins / Q000096:biosynthesis*; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010750:Phosphoproteins; D014764:Viral Proteins; C075740:P protein, Phocine distemper virus