CVI Accepts, published online ahead of print on 14 October 2009
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Clin. Vaccine Immunol. doi:10.1128/CVI.00255-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Characterization of an Antibody Depletion Assay for Analysis of Bactericidal Antibody Specificity

Wendell D. Zollinger*, Elizabeth E. Moran, and Deborah Schmiel

Walter Reed Army Institute of Research, 503 Robert Grant Ave., Silver Spring, MD, 20910-7500 USA

* To whom correspondence should be addressed. Email: wendell.zollinger{at}us.army.mil.


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Abstract

Serum bactericidal antibodies are important for protection against systemic Neisseria meningitidis infections. Consequently, identifying the specific targets of bactericidal antibodies is important for understanding protective immunity to meningococcal disease and for vaccine development and evaluation. We have developed a new assay that can be used to investigate the specificity of serum bactericidal antibodies. Prior to testing for bactericidal activity, antibodies specific for a given antigen or group of antigens are depleted from a serum by incubation with the antigen(s) bound to the wells of a 96-well microplate. A dilution series of the antigen is bound to the plate to assess the effectiveness of the antigen in removing the bactericidal antibodies. Removal of antibodies with solid phase antigen prior to bactericidal testing avoids depletion of complement by soluble immune complexes that can form when soluble antigen is present in the bactericidal test mixture (direct inhibition). The parameters associated with this assay are investigated and compared with those of a direct inhibition assay. The bactericidal depletion assay can be an effective tool for studying the specificity of serum bactericidal antibodies.






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