Differential reactivity of anti-primate and anti-human secondary antibodies against human and monkey immunoglobulins: implications for determining the sensitivity of immunogenicity assays

J Immunol Methods. 2005 Aug;303(1-2):76-80. doi: 10.1016/j.jim.2005.06.003.

Abstract

Development of immunogenicity assays for assessment of human antibodies to therapeutic proteins requires a quantitative determination of assay sensitivity. In the absence of true human positive controls, this is usually accomplished by utilizing affinity-purified antibodies from non-human primates or monoclonal antibodies. In the former case, it is generally considered that non-human primate antibodies will be recognized equally to human antibodies by secondary anti-human immunoglobulin reagents used in immunogenicity assays. We present results here demonstrating that this is not the case. In reality, anti-human immunoglobulin secondary antibodies do not recognize primate immunoglobulins as well as human immunoglobulins. As a result, the use of affinity purified primate antibodies to determine the sensitivity of an immunogenicity assay will likely result in the true sensitivity of the assay being underestimated.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Enzyme-Linked Immunosorbent Assay / standards
  • Enzyme-Linked Immunosorbent Assay / statistics & numerical data*
  • False Negative Reactions
  • Goats
  • Humans
  • Immunoglobulin A / metabolism
  • Immunoglobulin G / metabolism
  • Immunoglobulins / metabolism*
  • Macaca fascicularis*
  • Macaca mulatta*
  • Rabbits
  • Sensitivity and Specificity

Substances

  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulins