Characterization of antigens of Paragonimus miyazakii by ELISA.
The characterization of adult worm and excretory-secretory (ES) antigens of P. miyazakii was performed with a sandwich enzyme-linked immunosorbent assay (ELISA) IgG and Fab' fragments were prepared from P. miyazakii-infected rat sera. The former was purified by affinity column chromatography. The latter was prepared from IgG2c by passage through a Protein A affinity column, then conjugated to horseradish peroxidase. Cross-reactivity of the ELISA with Fasciola hepatica. Fischoederius elongatus. Taenia saginata. Schistosoma mansoni and S. Japonicum was negligible. With Paragonimus westermani, the same genus as P. miyazakii, cross-reactivity was below 30%. The molecular weight of both antigens was comparable (36,000) as determined by Sephadox G-100 gel filtration. Isoelectric points in both antigens had many peaks in a pH range of 3.9-4.7. In adult worm, pI of the major peaks were 4.6 and 4.7 and minor part of peaks had the common isoelectric points with four ES peaks in a pH range of 4.1-4.6. Antigenicity per protein concentration of both antigens was examined by the ELISA. The ES antigen was about five times as active antigenically as adult worm antigen. These results suggest that ES is one of the major antigens inducing antibody synthesis in rats infected with P. miyazakii, and that other antigens may be present within the adult worm.
Takemoto
Y
Y
Department of Parasitology, School of Medicine, University of Hiroshima, Japan.
Tsuji
M
M
D000818:Animals; D000947:Antigens, Helminth / Q000302:isolation & purification*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D010238:Paragonimus / Q000276:immunology*; D051381:Rats
D000947:Antigens, Helminth