Canine IgE monoclonal antibody specific for a filarial antigen: production by a canine x murine heterohybridoma using B cells from a clinically affected lymph node.

摘要: Canine popliteal lymph node cells taken at the onset of clinical disease from a rear limb infected with filarial nematode Brugia pahangi were fused mouse myeloma cell line P3X63.Ag8.653 cells. Of several canine immunoglobulin-producing clones this fusion, one was found to produce IgE specific for antigen. The has undergone limiting dilution cloning six times over past 3 years and continues monoclonal antibody subclass rate greater than mg/l. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) culture supernatant protein that bound A beads, showed bands molecular weights (MW) approximately 75,000 25,000 characteristic epsilon kappa or lambda chains, respectively. MW band, as demonstrated by Western blot. blots aqueous extracts adult nematodes binding single 35,000 peptide B. but not Dirofilaria immitis; immunochemistry using frozen sections worms, microfilariae fourth stage larvae revealed focal worm tissue adjacent dorsal ventral cords only adults.