[Preparation of monoclonal antibodies against Trichinella spiralis and localization of antigens].
BALB/c mice were infected per os with the infective larvae (L1) of Trichinella spiralis, and challenged by injecting 0.2ml soluble L1 antigen 3 days before cell fusion. SP2/0 myeloma and immune spleen cells were fused at the ratio of 1:5 in the presence 30%PEG (MW 4,000). The fusion rate and antibody positive rate were 92.1% and 16.6% respectively using ELISA. Among these the chromosomes of 4 strains were 2n greater than 100 and that of the SP2/0 myeloma cells 2n less than or equal to 70. The titer of McAb in the ascites was found to be 1/640 or 1/1,280. Eight strains were of IgG1, 1 IgG2a and 3 IgM in an agar system. Three strains of McAbs, which could recognize specifically the L1 antigen fractions by immuno-blotting technique, were used as probes to localize the target antigens in sections of T. spiralis L1. Staining was performed using an indirect technique consisting of goat anti-mouse IgG-conjugated horseradish peroxidase on de-paraffin sections, and substrate DAB. The results revealed that the antigens reacted most strongly with the specific McAbs were located in the cuticle and stichosome, followed by the lining of gut. Among the 3 strains of McAbs used, 2E5F11A5 reacted most strongly with the antigen, followed by 2E5E9E4. 2E5E9G5 was the weakest.
Feng
R
R
Yan
H
H
Liu
Y
Y
D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000947:Antigens, Helminth / Q000032:analysis*; D002459:Cell Fusion; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D014234:Trichinella / Q000276:immunology*
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth