Garbage in, garbage out: a critical evaluation of strategies used for validation of immunohistochemical biomarkers

Mol Oncol. 2014 Jun;8(4):783-98. doi: 10.1016/j.molonc.2014.03.008. Epub 2014 Mar 21.


The use of immunohistochemistry (IHC) in clinical cohorts is of paramount importance in determining the utility of a biomarker in clinical practice. A major bottleneck in translating a biomarker from bench-to-bedside is the lack of well characterized, specific antibodies suitable for IHC. Despite the widespread use of IHC as a biomarker validation tool, no universally accepted standardization guidelines have been developed to determine the applicability of particular antibodies for IHC prior to its use. In this review, we discuss the technical challenges faced by the use of immunohistochemical biomarkers and rigorously explore classical and emerging antibody validation technologies. Based on our review of these technologies, we provide strict criteria for the pragmatic validation of antibodies for use in immunohistochemical assays.

Keywords: Antibody reliability; Antibody validation; Biomarker discovery; Immunohistochemistry; Workflow.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Antibodies / metabolism
  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / metabolism*
  • Humans
  • Immunohistochemistry / methods*
  • Immunohistochemistry / standards
  • Neoplasms / diagnosis
  • Neoplasms / metabolism
  • Reference Standards
  • Specimen Handling / methods
  • Specimen Handling / standards
  • United States
  • United States Food and Drug Administration
  • Validation Studies as Topic


  • Antibodies
  • Biomarkers, Tumor