Production of a mouse monoclonal antibody against the alkaline phosphatase of adult Schistosoma mansoni.
This study describes the production and characterization of a monoclonal antibody (mAb) against the alkaline phosphatase of Schistosoma mansoni from splenocytes of chronically infected mice. Convenient selection of the mAb was achieved using the catalytic activity of the antigen in a developed enzyme-antigen immunoassay. The mAb was of the IgG1 subclass and it specifically recognized the alkaline phosphatase in adult worm sections by indirect immunofluorescence. Preincubation of the antibody with partially purified adult alkaline phosphatase did not result in inhibition of the enzyme activity and it did not mediate complement-dependent cytotoxicity against mechanically transformed schistosomula in vitro. The mAb was able to immunoprecipitate under reducing conditions a polypeptide of 65 kDa, similar in size to the monomeric subunit of the schistosome enzyme. The specificity of the mAb was assessed by competitive inhibition with antibodies of infected human sera in an immunoadsorption assay. Periodate treatment of the antigen resulted in altered electrophoretic mobility of alkaline phosphatase, which confirmed the presence of carbohydrate in the molecule, but this did not prevent binding by the mAb. Although the use of the mAb in capture assays for detection of circulating alkaline phosphatase in infected host sera was unsuccessful, the production of this antibody confirmed that the enzyme is exposed by adult worms to the host and that it is immunogenic; additionally, a monoclonal probe is available for further characterization of the structure and function of this important parasite surface molecule.
Pujol
F H
FH
Immunoparasitology Laboratory, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela.
Liprandi
F
F
Rodríguez
M
M
Cesari
I M
IM
D000469:Alkaline Phosphatase / Q000097:blood / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007163:Immunosorbent Techniques; D051379:Mice; D010504:Periodic Acid / Q000494:pharmacology; D011233:Precipitin Tests; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000276:immunology; D012555:Schistosomiasis mansoni / Q000276:immunology
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D007074:Immunoglobulin G; D010504:Periodic Acid; C009288:metaperiodate; D000469:Alkaline Phosphatase