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Detection of biomolecular interactions using polyfluorine tracers in diffusion ordered spectroscopy.

Tasneem S Habib¹, Abdulrahman M Alazemi¹, Laila A Alhaddad¹

¹Chemistry Department, Faculty of Science, Kuwait University, P.O. Box 5969, Safat 13060, Kuwait. a.alazmi@ku.edu.kw.

Analytical Methods : Advancing Methods and Applications

|November 17, 2025

Summary

This study introduces Diffusion Ordered Spectroscopy (DOSY) with a polyfluorine tracer to detect molecular interactions, successfully confirming avidin-biotin binding by measuring diffusion coefficients and determining structural parameters.

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Area of Science:

Analytical Chemistry
Biophysical Chemistry
Biochemistry

Background:

Biosensor technology faces challenges in detecting low-concentration protein complexes and understanding protein interactions.
Existing methods may require secondary antibodies or extensive purification, complicating analysis.

Purpose of the Study:

To develop an alternative biosensing method using Diffusion Ordered Spectroscopy (DOSY) to detect molecular interactions.
To investigate the avidin-biotin interaction using a polyfluorine tracer and DOSY.

Main Methods:

Utilized Diffusion Ordered Spectroscopy (DOSY) to measure diffusion coefficients of molecules before and after interaction.
Employed a pentadecafluorooctanoyl tracer to investigate the avidin-biotin interaction.
Calculated diffusion coefficients and hydrodynamic radius of the complex.

Main Results:

Confirmed avidin-biotin interaction by observing a significant decrease in diffusion coefficient upon binding (1.937 × 10^-8 m² s^-1 unbound vs. 1.12 × 10^-11 m² s^-1 bound).
Determined the hydrodynamic radius of the avidin-biotin-pentadecafluorooctanoyl complex to be 23 Å, consistent with literature values for native avidin.
Demonstrated accurate and specific results without secondary antibodies or purification.

Conclusions:

DOSY with polyfluorine tracers is an effective method for detecting molecular interactions and determining structural parameters.
This technique offers a simplified approach for biosensing applications in healthcare and military sectors.
The method provides a robust alternative for analyzing protein-protein interactions and molecular assemblies.