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Indirect enzyme-linked method for determining biotin in human serum

J O Nyalala¹, E Livaniou, L Leondiadis

¹Radioimmunochemistry Lab., NCSR Demokritos, Athens, Greece.

Journal of Immunoassay

|February 1, 1997

Summary

A new indirect enzyme-linked assay accurately quantifies biotin levels in human serum. This method offers a sensitive and reliable tool for measuring biotin concentrations across diverse populations.

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Area of Science:

Biochemistry
Analytical Chemistry
Clinical Diagnostics

Background:

Biotin, a vital B vitamin, plays a crucial role in various metabolic processes.
Accurate quantification of biotin in human serum is essential for diagnosing deficiencies and monitoring health status.

Purpose of the Study:

To develop and validate an indirect enzyme-linked assay for quantifying biotin concentrations in human serum.
To establish the assay's performance characteristics, including sensitivity, specificity, and precision.

Main Methods:

An indirect enzyme-linked assay was developed using streptavidin-conjugated horseradish peroxidase (streptavidin-HRP) and biotinylated bovine IgG (B-IgGb).
Biotin standards and unknown samples were preincubated with streptavidin-HRP before addition to coated plates.

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Main Results:

The assay demonstrated a dynamic range of 5 to 640 ng/L with a detection limit of 2 ng/L.
Intra- and inter-assay coefficients of variation (C.V.) were 1.6-3.9% and 3.7-7.2%, respectively.
Serum biotin levels were determined for healthy individuals (66-600 ng/L), pregnant women (60-360 ng/L), and hemodialysis patients (0.56-1.62 µg/L).

Conclusions:

The developed indirect enzyme-linked assay is a sensitive and reliable method for biotin quantification in human serum.
This assay provides valuable data for assessing biotin status in various physiological and pathological conditions.