title: Human hybridoma generation by hypo-osmolar electrofusion: characterization of human monoclonal antibodies to Schistosoma mansoni parasite antigens.
abstract: Human monoclonal antibodies which bind Schistosoma mansoni worm and egg antigens were identified and characterized from hybridomas generated using the hypo-osmolar electrofusion technique of somatic cell fusion. Splenocytes from S. mansoni infected individuals were mitogen-activated in vitro and subsequently fused by electrofusion. The greatest number of HAT resistant hybridomas per helical fusion chamber was obtained with unfrozen splenocytes cultured for 4-6 days after introduction of mitogen. Hybridomas secreting IgG antibodies recognizing parasite antigens were identified by ELISA. Twenty-one cloned cell lines secreting IgG antibody were maintained for at least 6 months. Characterization of antigen reactivity by Western blot analysis of nien cloned cell lines revealed antibodies which bound stage specific parasitic antigens. The data show that the technique of hypo-osmolar electrofusion produces stable, antibody producing hybridomas. The human monoclonal antibodies screened represent candidate molecules useful in the investigations of the human pathogen S. mansoni.
authors.affiliation: Department of Medicine, Miriam Hospital, Brown University, Providence.
authors.forename: G
authors.initials: G
authors.lastname: Klock
authors.forename: A V
authors.initials: AV
authors.lastname: Wisnewski
authors.forename: E A
authors.initials: EA
authors.lastname: el-Bassiouni
authors.forename: M I
authors.initials: MI
authors.lastname: Ramadan
authors.forename: P
authors.initials: P
authors.lastname: Gessner
authors.forename: U
authors.initials: U
authors.lastname: Zimmermann
authors.forename: T F
authors.initials: TF
authors.lastname: Kresina
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000737:chemistry; D002459:Cell Fusion; D004560:Electricity; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G; D008970:Molecular Weight; D009994:Osmolar Concentration; D012550:Schistosoma mansoni / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D007074:Immunoglobulin G