Search for epitopic sites of antibodies to germ cell alkaline phosphatase.
This study comprises a search for epitopic sites of the germ cell alkaline phosphatase by the use of polyclonal and monoclonal antibodies. The aim of this study was to define epitopes of the germ cell alkaline phosphatase molecule by synthetic peptides representing the known sequence of the molecule. The amino acid sequences of the epitopes to which antisera react were not known. We used overlapping peptides covering the whole 532 amino acid germ cell alkaline phosphatase molecule, including the signal 19 amino acid peptide. A polyclonal, and three monoclonal, antibodies known to react equally well with placental alkaline phosphatase isozyme (PLAP), but which differ in their reactivities with the germ cell alkaline phosphatase isozyme, were used. We have revealed potential epitopic sites of the germ cell alkaline phosphatase molecule using first polyclonal anti-PLAP, followed by monoclonal antibodies for the mapping. We could also show that the antibodies exhibited higher reactivities with such sites if they were combined in a linear synthesis. This indicates that the germ cell alkaline phosphatase molecule has discontinuous epitopes.
Harmenberg
U
U
Department of Virology, National Bacteriological Laboratory, Stockholm, Sweden.
Ljungdahl-Ståhle
E
E
Rudén
U
U
Wahren
B
B
D000469:Alkaline Phosphatase / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D000906:Antibodies; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005854:Germ Cells / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007527:Isoenzymes; D009842:Oligopeptides; D000469:Alkaline Phosphatase