Targeting the Antibody Fab Region Using Light-Induced Indole-3-Butyric Acid Functionalized Magnetic Microspheres

Area of Science:

• Biotechnology
• Materials Science
• Biochemistry

Background:

• Antibody fragments (Fab) are crucial in diagnostics and therapeutics.
• Current purification methods often lack specificity and efficiency.
• Developing targeted and adjustable purification systems is essential.

Purpose of the Study:

• To develop a novel light-controlled adsorption system for direct targeting of antibody Fab fragments.
• To functionalize magnetic microspheres with indole-3-butyric acid for specific ligand conjugation.
• To evaluate the system's adsorption, kinetics, selectivity, and reusability.

Main Methods:

• Conjugation of indole-3-butyric acid to amine-functionalized magnetic microspheres using EDC/NHS chemistry.
• Light-induced radical generation for antibody Fab fragment adsorption.
• Static adsorption and kinetic studies to analyze adsorption behavior (Langmuir model, second-order kinetics).
• Evaluation of elution conditions, recovery rates, and reusability.

Main Results:

• The system demonstrated specific targeting and effective affinity adsorption of antibody Fab fragments.
• Adsorption followed the Langmuir model (K_F = 0.122, R² = 0.996) and second-order kinetics (k₂ = 0.0257, R² = 0.989).
• High purification recovery (>98%) and antigen-binding activity retention were achieved.
• The system showed excellent regenerability, retaining >96% efficiency after 10 reuse cycles.

Conclusions:

• A highly efficient, stable, and cost-effective light-controlled adsorption system for antibody Fab purification was successfully developed.
• The system offers enhanced selectivity, adjustable properties, and excellent reusability.
• This technology presents a promising future method for antibody Fab purification.