Surface plasmon resonance measurements of plasma antibody avidity during primary and secondary responses to anthrax protective antigen

J Immunol Methods. 2014 Feb;404:1-12. doi: 10.1016/j.jim.2013.11.026. Epub 2013 Dec 4.


Establishment of humoral immunity against pathogens is dependent on events that occur in the germinal center and the subsequent induction of high-affinity neutralizing antibodies. Quantitative assays that allow monitoring of affinity maturation and duration of antibody responses can provide useful information regarding the efficacy of vaccines and adjuvants. Using an anthrax protective antigen (rPA) and alum model antigen/adjuvant system, we describe a methodology for monitoring antigen-specific serum antibody concentration and avidity by surface plasmon resonance during primary and secondary immune responses. Our analyses showed that following a priming dose in mice, rPA-specific antibody concentration and avidity increases over time and reaches a maximal response in about six weeks, but gradually declines in the absence of antigenic boost. Germinal center reactions were observed early with maximal development achieved during the primary response, which coincided with peak antibody avidity responses to primary immunization. Boosting with antigen resulted in a rapid increase in rPA-specific antibody concentration and five-fold increase in avidity, which was not dependent on sustained GC development. The described methodology couples surface plasmon resonance-based plasma avidity measurements with germinal center analysis and provides a novel way to monitor humoral responses that can play a role in facilitating vaccine and adjuvant development.

Keywords: Antibody avidity; Germinal centers; Protective antigen (PA); Surface plasmon resonance (SPR).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adjuvants, Immunologic / administration & dosage
  • Alum Compounds / administration & dosage
  • Animals
  • Anthrax / blood
  • Anthrax / immunology
  • Anthrax / microbiology
  • Anthrax / prevention & control*
  • Anthrax Vaccines / administration & dosage
  • Anthrax Vaccines / immunology*
  • Antibodies, Bacterial / blood*
  • Antibodies, Neutralizing / blood*
  • Antibody Affinity
  • Antibody Specificity
  • Antigens, Bacterial / immunology
  • Bacillus anthracis / immunology*
  • Bacillus anthracis / pathogenicity
  • Bacterial Toxins / antagonists & inhibitors*
  • Bacterial Toxins / immunology
  • Female
  • Germinal Center
  • Immunity, Humoral*
  • Immunization, Secondary
  • Mice
  • Mice, Inbred C57BL
  • Surface Plasmon Resonance


  • Adjuvants, Immunologic
  • Alum Compounds
  • Anthrax Vaccines
  • Antibodies, Bacterial
  • Antibodies, Neutralizing
  • Antigens, Bacterial
  • Bacterial Toxins
  • anthrax toxin
  • aluminum sulfate