Human hybridoma generation by hypo-osmolar electrofusion: characterization of human monoclonal antibodies to Schistosoma mansoni parasite antigens.
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.
Klock
G
G
Department of Medicine, Miriam Hospital, Brown University, Providence.
Wisnewski
A V
AV
el-Bassiouni
E A
EA
Ramadan
M I
MI
Gessner
P
P
Zimmermann
U
U
Kresina
T F
TF
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*
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D007074:Immunoglobulin G