Isolation of Trichinella-specific antigens for diagnosis by gradient monoclonal antibody affinity chromatography

Monoclonal antibodies were generated for the isolation of specific antigens from Trichinella spiralis. Two monoclonal antibodies (7G6-2 and 10B6-1) of class IgG2b and IgG1 were selected according to their reactivities in an enzyme-linked immunosorbent assay and western blot. Clone 7G6-2 reacted with an antigen with molecular mass of approximately 60 kDa, and clone 10B6-1 bound to multiple antigens ranging from 49 to 62 kDa on western blot. Antibodies of each clone were purified partially from mouse ascites fluid by ammonium sulfate precipitation and were coupled to CNBr-activated Sepharose 4B. Antigens with molecular masses of 49 kDa and 57 kDa (P49/57), 52-62 kDa (P52/62), and 60 kDa (P60) were isolated from larval excretory-secretory products and crude worm extract with column 10B6-1 and column 7G6-2, respectively, in part by changing the pH of elution buffers. These antigens were mostly glycoproteins, strongly immunogenic, and specific to the parasite.