Using liposomal fluorescent biolabels to develop an immunoaffinity chromatographic biosensing system for biotin

Area of Science:

• Analytical Chemistry
• Biotechnology
• Biochemistry

Background:

• Biotin is essential for cellular function and growth.
• Current analytical methods for biotin quantitation lack speed, simplicity, and reliability.
• Accurate biotin measurement is crucial for bodily fluids and foodstuffs analysis.

Purpose of the Study:

• To develop a rapid, simple, and reliable analytical tool for direct biotin determination.
• To establish an immunoaffinity chromatographic biosensing system for biotin quantitation.
• To enable sensitive and accurate measurement of biotin in diverse sample matrices.

Main Methods:

• Synthesis of a stationary phase by immobilizing antibiotin monoclonal antibodies onto NHS-activated sepharose beads.
• Development of a competitive assay using biotin-tagged liposomes encapsulating carboxyfluorescein (CF) as biolabels.
• Quantification of biotin by measuring fluorescence intensity of released CF after methanol-induced liposome lysis.

Main Results:

• The developed immunoaffinity chromatographic biosensing system allows direct determination of biotin.
• The assay demonstrated a linear calibration curve for biotin over 8 orders of magnitude (10^-12 to 10^-4 M).
• A low limit of detection of 5.0 pg of biotin was achieved.

Conclusions:

• The proposed immunoaffinity chromatographic biosensing system is a sensitive and reliable method for biotin quantitation.
• This biosensing system overcomes limitations of existing analytical tools for biotin measurement.
• The method is suitable for direct determination of biotin in bodily fluids and foodstuffs.