Two-dimensional electrophoresis and mass spectrometry for the identification of species-specific Trichinella antigens.
A proteomic approach was utilized for fine antigenic characterization of the closely related Trichinella genotypes Trichinella britovi T3 and Trichinella T8. Crude extract of muscle larvae L1 (LCE) from both isolates were analyzed by 2D-PAGE. Over 500 protein spots were reproducibly separated in both genotypes. These separated proteins were identified in Western blot with IgG1 and IgG3 from homologous and heterologous hyperimmune sera raised in BALB/c mice. A group of 20 and 15 spots migrating at 50--60 k Da and pH 5.5--6.5 in T. britovi and Trichinella T8 maps, respectively, reacted with the IgG1 from heterologous sera whereas a group of minor spots of similar migration patterns did not. Low cross-reactivity occurred for IgG3. MALDI-TOF and MALDI-TOF/TOF analysis of these antigens identified a predicted enolase, the protein P 49 and a predicted actin among the cross-reactive proteins and two hypothetical actins among the non cross-reactive proteins.
Department of Parasitology, Faculty of Pharmacy, Complutense University, Plaza Ramón y Cajal s/n, Ciudad Universitaria 28040 Madrid, Spain.
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Dea-Ayuela
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Bolás-Fernández
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis* / Q000737:chemistry; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D007074:Immunoglobulin G / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D010449:Peptide Mapping / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014234:Trichinella / Q000276:immunology*
D000947:Antigens, Helminth; D007074:Immunoglobulin G