Depletion of biotin using streptavidin-coated microparticles: a validated solution to the problem of biotin interference in streptavidin-biotin immunoassays

Ann Clin Biochem. 2018 Mar;55(2):216-226. doi: 10.1177/0004563217707783. Epub 2017 Jun 29.


Background Biotin interference in streptavidin-based immunoassays causes widespread analytical distortions that may lead to clinical confusion, inappropriate patient management and, ultimately, adverse events. Its prevalence has increased recently due to the increased use of high-dose biotin therapy in specific patient groups (notably multiple sclerosis) and possibly the general community. Methods We have developed a method to deplete biotin from samples using the streptavidin-coated magnetic microparticles that are a component of most susceptible assays. Results We show that high concentrations of spiked biotin can be adequately depleted from serum using this approach, and that gross biochemical derangements can be restored to normality. We also show that biotin in samples derived from multiple sclerosis patients receiving 300 mg biotin daily can be adequately depleted to remove associated analytical interference and restore normal results. The method is applicable to competitive and sandwich immunoassays and importantly, because it does not change the volume of the sample, suitable for the measurement of free thyroid hormone assays. Application of the method does not significantly change the precision of measurement, and for the majority of analytes, the accuracy is not substantially altered. Conclusions Adopting this method enables laboratories to confirm biotin interference in the appropriate clinical setting. Moreover, it enables laboratories to remove the interference and report accurate and reliable results, without the need for patients to withhold beneficial therapy prior to blood tests. Until the biotin tolerance of susceptible assays is improved, our method gives laboratories a safe alternative for reporting results using streptavidin-based methods.

Keywords: Evaluation of new methods; immunoassay; laboratory methods.

Publication types

  • Validation Study

MeSH terms

  • Artifacts
  • Biotin / blood
  • Biotin / chemistry
  • Biotin / isolation & purification*
  • Biotin / therapeutic use
  • Clinical Laboratory Techniques / methods
  • Clinical Laboratory Techniques / standards
  • Coated Materials, Biocompatible / chemistry*
  • Humans
  • Immunoassay / methods*
  • Immunoassay / standards
  • Multiple Sclerosis / drug therapy
  • Multiple Sclerosis / physiopathology
  • Streptavidin / chemistry*
  • Thyroid Function Tests


  • Coated Materials, Biocompatible
  • Biotin
  • Streptavidin