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Synergetic collision and space separation in microfluidic chip for efficient affinity-discriminated molecular

Junxia Wang^1,2, Liang Li¹, Yingkun Zhang¹

¹The MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.

Proceedings of the National Academy of Sciences of the United States of America

|October 3, 2022

Summary

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This study introduces a microfluidic system to enhance molecular selection efficiency. The novel approach improves the isolation of high-affinity ligands for biomedical applications.

Area of Science:

Biotechnology
Microfluidics
Molecular Biology

Background:

Efficient molecular selection is crucial for developing diagnostic, therapeutic, and vaccinology tools.
Selection efficiency is heavily influenced by the dissociation equilibrium achievable in a single round.
Shifting equilibrium towards dissociation favors high-affinity ligands, enhancing selection.

Purpose of the Study:

To synergize dual effects using deterministic lateral-displacement microfluidics to significantly shift equilibrium.
To improve affinity discrimination in molecular selection processes.
To demonstrate a novel strategy for discovering robust binding ligands.

Main Methods:

Utilized deterministic lateral-displacement microfluidics, incorporating collision-based force and 2D separation-based concentration effects.

Keywords:

affinity discrimination microfluidics molecular selection phage display

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Applied the system to phage display for both experimental and simulation-based validation.

Tested the system's efficacy in selecting high-affinity ligands against tumor markers EphA2 and CD71.

Main Results:

The microfluidic system effectively shifted the dissociation equilibrium, enhancing selection efficiency.
Demonstrated improved removal of low- and moderate-affinity ligands while retaining high-affinity ones.
Successfully obtained two high-affinity and specific peptides targeting EphA2 and CD71 within a single selection round.

Conclusions:

The proposed microfluidic strategy significantly improves affinity discrimination in molecular selection.
This approach offers a promising direction for the discovery of high-affinity ligands for diverse biomedical applications.
The system enhances the generation of molecular tools for diagnosis, pathology, vaccinology, and therapeutics.