Microfluidic on-chip capture-cycloaddition reaction to reversibly immobilize small molecules or multi-component...

Area of Science:

• Bioconjugation Chemistry
• Surface Science
• Biosensor Technology

Background:

• Developing efficient methods for surface immobilization of bioactive molecules is crucial for advancing biosensor and microarray technologies.
• Controlling molecular orientation and immobilization density is essential for optimizing sensor performance.

Purpose of the Study:

• To establish a microfluidic method for rapid and controlled surface immobilization of small molecules using bioorthogonal chemistry.
• To enable real-time monitoring of immobilization processes and facilitate reversible capture for extended experimental applications.

Main Methods:

• Utilized a covalent bioorthogonal [4+2] cycloaddition reaction between trans-cyclooctene (TCO) and 1,2,4,5-tetrazine (Tz) for molecule immobilization.
• Employed surface plasmon resonance (SPR) for real-time monitoring under continuous flow conditions.
• Integrated non-covalent antigen-antibody capture with cycloaddition for reversible immobilization and multi-component assembly.

Main Results:

• Demonstrated successful microfluidic immobilization of TCO/Tz-derivatized molecules with real-time SPR monitoring.
• Achieved reversible immobilization by combining cycloaddition with antigen-antibody interactions.
• Showcased the method's versatility through immobilization of a small molecule (AP1497) and a functionalized nanoparticle on a biosensor chip.

Conclusions:

• The developed method provides a versatile platform for on-chip assembly and interaction studies of multi-component structures.
• This approach enhances biosensor capabilities by allowing for controlled, reversible, and real-time immobilization of diverse molecules.
• The technique holds significant potential for applications in diagnostics, drug discovery, and fundamental biological research.