A Biomimetic Protein Immobilization Method for Studying Drug-Protein Interactions Based on Affinity Capillary...

Area of Science:

• Analytical Chemistry
• Biomaterials Science
• Pharmacology

Background:

• Accurate determination of drug-protein interaction dissociation constants (Kd) is crucial but lacks a universal method.
• Traditional affinity capillary electrophoresis (ACE) faces limitations in complex systems.

Purpose of the Study:

• To develop a novel biomimetic affinity capillary electrochromatography (ACEC) platform for determining drug-protein interactions (DPI).
• To establish a universal tool for high-throughput drug screening and structure-guided therapeutic design.

Main Methods:

• Fabrication of a polydopamine (PDA) and human serum albumin (HSA) coated capillary column (PDA/PDA/HSA@capillary).
• Optimization of PDA coating (0.8 mg/mL) and HSA immobilization (0.25 mM) concentrations.
• Characterization of the capillary column and evaluation of its separation performance in complex drug systems.

Main Results:

• Successful coating of PDA and immobilization of HSA on the capillary inner wall.
• Demonstrated good separation effects for various drugs and accurate Kd values for rutin, quercitrin, and quercetin.
• Consistent results when applied to Sophora japonica extract and high reproducibility over 50 runs.

Conclusions:

• The developed PDA/PDA/HSA@capillary platform provides a universal and effective method for determining Kd values in DPI.
• This biomimetic ACEC approach overcomes critical bottlenecks in traditional ACE, enabling efficient drug screening.
• The study offers a valuable tool for advancing structure-guided therapeutic design.