Two-dimensional electrophoresis and mass spectrometry for the identification of species-specific Trichinella antigens

• Published in: Veterinary parasitology Vol. 132; no. 1; pp. 43 - 49
• Main Authors: Dea-Ayuela, M.A., Bolás-Fernández, F.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05-09-2005
• Subjects: 2D-electrophoresis; Amino Acid Sequence; Animals; antigen-antibody reactions; Antigens; Antigens, Helminth - analysis; Antigens, Helminth - chemistry; Blotting, Western; cross reaction; Electrophoresis, Gel, Two-Dimensional; immunoblotting; immunoglobulin G; Immunoglobulin G - analysis; MALDI-TOF mass spectrometry; Mass spectrometry; Mice; Mice, Inbred BALB C; Peptide Mapping - methods; polyacrylamide gel electrophoresis; proteomics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Trichinella; Trichinella - immunology; Trichinella britovi; Western blotting; 2D-electrophoresis; Antigens; Mass spectrometry; Trichinella
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/j.vetpar.2005.05.018

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
kDa 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 P49 and a predicted actin among the cross-reactive proteins and two hypothetical actins among the non cross-reactive proteins.