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Surface Plasmon Resonance Analysis of Antipolysaccharide Antibody Specificity: Responses to Meningococcal Group C Conjugate Vaccines and Bacteria

Pablo A. García-Ojeda,¹ Sharon Hardy,² Steven Kozlowski,¹ Kathryn E. Stein,^1, and Ian M. Feavers^2*

Division of Monoclonal Antibodies, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892,¹ Division of Bacteriology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Hertfordshire, England EN6 3QG²

Received 11 November 2003/ Returned for modification 23 December 2003/ Accepted 19 February 2004

Antibody (Ab) responses to polysaccharides (PS), such as Neisseria meningitidis group C PS (MCPS), are characterized as being thymus independent and are restricted with regard to clonotype and isotype expression. PS conjugated to proteins, e.g., MCPS coupled with tetanus toxoid or the diphtheria toxin derivative CRM₁₉₇, elicit thymus-dependent responses. The present study developed a surface plasmon resonance approach to evaluate Ab responses to MCPS conjugate vaccines, including either O-acetylated (OAc⁺) or de-O-acetylated (OAc^–) forms of the PS. The results were generally consistent with those obtained by enzyme-linked immunosorbent assay and showed that sera from mice immunized with conjugate vaccines contain Abs that bind more effectively to OAc⁺ and OAc^– MCPS than sera from mice immunized with fixed bacteria. The data suggest a critical shared or overlapping epitope recognized by all the conjugate vaccine immune sera and strategies for assessing polyclonal Ab avidity.

Editor: J. N. Weiser

Present address: MacroGenics, Inc., Rockville, MD 20850.

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