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Summary
Biocytin is typically undetectable in healthy individuals due to biotinidase activity. This study presents a new assay to detect biocytin in biological samples, even with high biotin levels, aiding diagnosis of biotinidase deficiency.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Analytical Chemistry
Background:
- Biocytin (epsilon-N-[d-biotinyl]-L-lysine) is usually absent in healthy individuals' bodily fluids because of widespread biotinidase enzyme activity.
- Elevated biocytin levels in serum and urine are hypothesized for individuals with inherited biotinidase deficiency.
Purpose of the Study:
- To develop a novel noncompetitive assay for quantifying biocytin in biological samples.
- To enable biocytin determination in samples containing both biotin and biocytin.
Main Methods:
- Utilized the specific interaction between biocytin and avidin for assay development.
- Employed an anion-exchange resin to remove interfering biotin from samples prior to assay.
- Validated a noncompetitive avidin-binding assay for accurate biocytin measurement.
Main Results:
- Successfully developed and validated a noncompetitive assay for biocytin.
- The assay effectively quantifies biocytin in the presence of biotin by removing biotin interference.
- The assay's performance is discussed concerning potential interference from elevated biotin levels.
Conclusions:
- The developed avidin-binding assay provides a reliable method for biocytin detection.
- This assay is valuable for diagnosing inherited biotinidase deficiency by measuring biocytin levels.
- The method addresses the challenge of co-existing biotin and biocytin in biological samples.