Quantification of biotin in serum by competition with solid-phase biotin for binding to peroxidase-avidin conjugate

We have developed a simple, fast, accurate, and reproducible method--an Enzyme-Linked Ligand Sorbent Assay (ELLSA)--for quantifying biotin without the use of isotopes and immunological reagents. The analytical principle is based on the competition between biotin of the sample and immobilized biotin, i.e., biotinylated bovine serum albumin coated onto immunoplates, for binding to horseradish peroxidase (EC 1.11.1.7)-avidin conjugate. The amount of conjugate bound to immobilized biotin is subsequently measured.