Immunoassay design and biotin interference

Biotin (vitamin B₇ or vitamin H), a member of vitamin B complex, acts as a cofactor for five biotin-dependent carboxylases, thus playing critical roles in gluconeogenesis, fatty acid synthesis and amino acid catabolism. Although rare inborn errors of metabolism may cause biotin deficiency, these can be successfully treated with biotin supplementation. In general, normal individuals do not get any benefit from taking biotin supplement. Nevertheless, biotin use remains widespread for growing healthy hair and nail. Unfortunately, the use/overuse of supplemental biotin may interfere with immunoassays that incorporate biotinylated antibody in assay design. Biotin if present in elevated concentration in serum or plasma, may falsely increase analyte concentration using competitive immunoassay (positive interference). In contrast, biotin shows negative interference if sandwich immunoassay format is used. Such interferences may cause diagnostic error, most commonly in cases of hyperthyroidism due to (1) positive interference of biotin in free triiodothyronine (FT3) and free thyroxine (FT4) assays (competitive format) and (2) negative interference in thyroid stimulating hormone (TSH) assay (sandwich format). In this review, I explore the biochemistry of biotin and discuss its role as a potential interferent in immunoassay formats that are biotin based.