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Journal Article | Research Support, Non-U.S. Gov't

Characterization of two conformational epitopes of the Chlamydia trachomatis serovar L2 DnaK immunogen.

S Birkelund, P Mygind, A Holm, B Larsen, F Beck, G Christiansen
S Birkelund
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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P Mygind
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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A Holm
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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B Larsen
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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F Beck
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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G Christiansen
Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark.
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ABSTRACT

Chlamydia trachomatis DnaK is an important immunogen in chlamydial infections. DnaK is composed of a conserved N-terminal ATP-binding domain and a variable C-terminal peptide-binding domain. To locate the immunogenic part of C. trachomatis Dnak, we generated monoclonal antibodies (MAbs) against this protein. By use of recombinant DNA techniques, we located the epitopes for two MAbs in the C-terminal variable part. Although the antibodies reacted in an immunoblot assay, it was not possible to map the epitopes completely by use of 16-mer synthetic peptides displaced by one amino acid corresponding to the C-terminal part of C. trachomatis DnaK. To determine the limits of the epitopes, C. trachomatis DnaK and glutatione S-transferase fusion proteins were constructed and affinity purified. The purified DnaK fusion proteins were used for a fluid-phase inhibition enzyme-linked immunosorbent assay with the two antibodies. The epitopes were found not to overlap. To obtain DnaK fragments recognized by the antibodies with the same affinity as native C. trachomatis DnaK, it was necessary to express, respectively, regions of 127 and 77 amino acids. The MAbs described in this study thus recognized conformational epitopes of C. trachomatis DnaK.

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Characterization of two conformational epitopes of the Chlamydia trachomatis serovar L2 DnaK immunogen.
S Birkelund, P Mygind, A Holm, B Larsen, F Beck, G Christiansen
Infection and Immunity Mar 1996, 64 (3) 810-817; DOI:

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Characterization of two conformational epitopes of the Chlamydia trachomatis serovar L2 DnaK immunogen.
S Birkelund, P Mygind, A Holm, B Larsen, F Beck, G Christiansen
Infection and Immunity Mar 1996, 64 (3) 810-817; DOI:
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