Document Detail

Validation of in silico prediction by in vitro immunoserological results of fine epitope mapping on citrate synthase specific autoantibodies.
MedLine Citation:
PMID:  16087237     Owner:  NLM     Status:  MEDLINE    
In silico antibody-antigen binding predictions are generally employed in research to rationalize epitope development. These techniques are widely spread despite their technical limitations. To validate the results of these bioinformatic calculations evidence based comparative in vitro studies are necessary. We have used a well-conserved mitochondrial inner membrane antigen-citrate synthase to develop a model for comparative analysis of the predicted and the immunoserologically verified epitopes of circulating autoantibodies. Epitopes were predicted using accepted tools: the GCG Wisconsin package and TEPITOPE 2000. An overlapping multipin ELISA assay--covering 49% of the citrate synthase molecule--was developed to map autoantibody epitopes of individuals (healthy, systemic autoimmune, and heart transplanted) in different immunopathological conditions. From the 40 synthesized decapeptides 34 were predicted in silico and 27 were validated in vitro. Thirty-two percent of epitopes were recognized by majority of sera 47% by at least one sera. False positive predictions were 21%. There was major difference in the recognized epitope pattern under different immunopathological conditions. Our results suggest that special databases are needed for training and weighing prediction methods by clinically well-characterized samples, due to the differences in the immune response under different health status. The development of these special algorithms needs a new approach. A high number of samples under these special immunological conditions are to be mapped and then used for the "fine tuning" of different prediction algorithms.
Z Nyárády; T Czömpöly; Sz Bosze; G Nagy; A Petrohai; J Pál; F Hudecz; T Berki; P Németh
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-08-08
Journal Detail:
Title:  Molecular immunology     Volume:  43     ISSN:  0161-5890     ISO Abbreviation:  Mol. Immunol.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2005-12-20     Completed Date:  2006-03-29     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7905289     Medline TA:  Mol Immunol     Country:  England    
Other Details:
Languages:  eng     Pagination:  830-8     Citation Subset:  IM    
Department of Immunology and Biotechnology, University of Pécs, Szigeti út 12, H-7643 Pécs, Faculty of Medicine, Hungary.
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MeSH Terms
Amino Acid Sequence
Antigen-Antibody Reactions / immunology*
Autoantibodies / chemistry*,  immunology
Cell Membrane / enzymology,  immunology
Citrate (si)-Synthase / immunology*
Computer Simulation
Epitope Mapping*
Epitopes, B-Lymphocyte / chemistry*,  immunology
Epitopes, T-Lymphocyte / chemistry*,  immunology
Models, Immunological
Molecular Sequence Data
Protein Conformation
Reg. No./Substance:
0/Autoantibodies; 0/Epitopes, B-Lymphocyte; 0/Epitopes, T-Lymphocyte; EC (si)-Synthase

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