Multiplexed Fc array for evaluation of antigen-specific antibody effector profiles

J Immunol Methods. 2017 Apr;443:33-44. doi: 10.1016/j.jim.2017.01.010. Epub 2017 Feb 3.


Antibodies are widely considered to be a frequent primary and often mechanistic correlate of protection of approved vaccines; thus evaluating the antibody response is of critical importance in attempting to understand and predict the efficacy of novel vaccine candidates. Historically, antibody responses have been analyzed by determining the titer of the humoral response using measurements such as an ELISA, neutralization, or agglutination assays. In the simplest case, sufficiently high titers of antibody against vaccine antigen(s) are sufficient to predict protection. However, antibody titer provides only a partial measure of antibody function, which is dependent on both the variable region (Fv) to bind the antigen target, and the constant region (Fc) to elicit an effector response from the innate arm of the immune system. In the case of some diseases, such as HIV, for which an effective vaccine has proven elusive, antibody effector function has been shown to be an important driver of monoclonal antibody therapy outcomes, of viral control in infected patients, and of vaccine-mediated protection in preclinical and clinical studies. We sought to establish a platform for the evaluation of the Fc domain characteristics of antigen-specific antibodies present in polyclonal samples in order to better develop insights into Fc receptor-mediated antibody effector activity, more fully understand how antibody responses may differ in association with disease progression and between subject groups, and differentiate protective from non-protective responses. To this end we have developed a high throughput biophysical platform capable of simultaneously evaluating many dimensions of the antibody effector response.

Keywords: ADCC; Antibody; Effector function; IgG; Phagocytosis; Vaccines.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • AIDS Vaccines / administration & dosage*
  • AIDS Vaccines / immunology
  • Animals
  • Antibody Affinity
  • Antibody-Dependent Cell Cytotoxicity
  • Binding Sites, Antibody
  • Case-Control Studies
  • Disease Models, Animal
  • Flow Cytometry
  • HIV / immunology*
  • HIV Antibodies / blood
  • HIV Antibodies / immunology*
  • HIV Infections / blood
  • HIV Infections / immunology
  • HIV Infections / therapy*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immunity, Humoral / drug effects*
  • Immunization
  • Immunoglobulin Fc Fragments / blood
  • Immunoglobulin Fc Fragments / immunology*
  • Immunologic Techniques*
  • Macaca mulatta
  • Phagocytosis
  • Protein Binding
  • Receptors, IgG / genetics
  • Receptors, IgG / immunology*
  • Receptors, IgG / metabolism
  • Simian Acquired Immunodeficiency Syndrome / blood
  • Simian Acquired Immunodeficiency Syndrome / immunology
  • Simian Acquired Immunodeficiency Syndrome / therapy
  • Simian Immunodeficiency Virus / immunology


  • AIDS Vaccines
  • HIV Antibodies
  • Immunoglobulin Fc Fragments
  • Receptors, IgG