NMR identification of epitopes of Lyme disease antigen OspA to monoclonal antibodies.
Outer surface protein A (OspA) from the Lyme disease spirochete Borrelia burgdorferi has been a focus of vaccine development. We have identified epitopes of OspA to two monoclonal antibodies (mAbs) by comparing NMR chemical shifts of free OspA and those in Fab complexes. Deuteration of non-labile protons in OspA extended the size limit of this technique so that it was applicable to the 78 kDa complexes of OspA and the Fab fragment. The epitope identified by NMR to an mAb, 184.1, agrees well with that previously defined by the crystal structure of the same complex, indicating the ability of the NMR method to accurately map an epitope in a large protein complex. The technique mapped the epitope to mAb 336, a mAb of clinical interest, to a region centered at the C-terminal alpha-helix. The results provides a basis for rational design of OspA-based Lyme disease vaccines.
Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, NY, 14642, USA.
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Luft
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D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000302:isolation & purification; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000276:immunology*; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D008074:Lipoproteins; D008193:Lyme Disease / Q000276:immunology* / Q000382:microbiology; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D008074:Lipoproteins; C072045:OspA protein