title: Detection of anti-PL-12 autoantibodies by ELISA using a recombinant antigen; study of the immunoreactive region.
abstract: Autoantibodies to aminoacyl-tRNA synthetases are highly associated with myositis and detection is important in clinical diagnosis; however, current methods of screening limit its clinical utility. In the present study, alanyl-tRNA synthetase (PL-12) recombinant protein was obtained by immunological screening of a HeLa expression library and used in an ELISA with 22 anti-PL-12 sera, 200 autoimmune sera negative for PL-12 and 100 healthy individual sera. Sensitivity of the method was 95% (21/22) and specificity 100%. Mapping of the immunoreactive region was carried out using three anti-PL-12 sera and different recombinant protein-derived peptides. Results show that the same conformational epitope located within amino acids 730-951 of the PL-12 antigen outside the catalytic region was recognized by the three anti-PL-12 sera tested. We conclude that ELISA using recombinant protein is an effective and useful method for routine screening for anti-PL-12 autoantibodies.
authors.affiliation: Servicio de Inmunología, Hospital Universitario Virgen del Rocío, Sevilla, Spain.
authors.forename: J R
authors.initials: JR
authors.lastname: García-Lozano
authors.forename: M F
authors.initials: MF
authors.lastname: González-Escribano
authors.forename: R
authors.initials: R
authors.lastname: Rodríguez
authors.forename: J L
authors.initials: JL
authors.lastname: Rodriguez-Sanchez
authors.forename: I N
authors.initials: IN
authors.lastname: Targoff
authors.forename: I
authors.initials: I
authors.lastname: Wichmann
authors.forename: A
authors.initials: A
authors.lastname: Núñez-Roldán
mesh_terms: D000412:Alanine-tRNA Ligase / Q000235:genetics / Q000276:immunology*; D001323:Autoantibodies / Q000032:analysis* / Q000276:immunology; D001324:Autoantigens / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D018985:Epitopes, B-Lymphocyte; D011994:Recombinant Proteins; D000412:Alanine-tRNA Ligase