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Enabling Flow-Based Kinetic Off-Rate Selections Using a Microfluidic Enrichment Device.

William E Evenson¹, Wan-Zhen Sophie Lin², Kenmond Pang²

¹Department of Chemistry, University of Southern California, 3620 McClintock Avenue, SGM 418, Los Angeles, California 90089, United States.

Analytical Chemistry

|July 8, 2020

Summary

Researchers developed a microfluidic enrichment device (MFED) for faster selection of high-affinity ligands. This method accelerates the study of diagnostic and therapeutic targets identified through genomic sequencing.

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Area of Science:

Biotechnology
Molecular Biology
Genomics

Background:

Genomic sequencing rapidly identifies potential diagnostic and therapeutic targets.
Existing methods struggle to produce the necessary peptide- and protein-based ligands for target study.
A bottleneck exists in ligand generation for novel targets.

Purpose of the Study:

To develop a novel method for kinetic off-rate selection of ligands.
To create a microfluidic enrichment device (MFED) that bypasses the need for exogenous competitors.
To accelerate the generation of peptide- and protein-based ligands for studying genomic targets.

Main Methods:

Development of a microfluidic enrichment device (MFED).
Kinetic off-rate selection without exogenous competitor.
Tuning device conditions (bed volume, flow rate, immobilized target) to control ligand binding kinetics.
Application of simple kinetic equations to model ligand binding.

Main Results:

The MFED enables kinetic off-rate selection by manipulating flow rates.
Observed kinetic rate constants align with independent measurements.
Demonstrated a 4-fold improvement in ligand enrichment compared to standard selection methods.
Reduced target-protein demand to less than a nanomole per selection.

Conclusions:

The MFED effectively biases ligand pools toward high-affinity binders.
This device accelerates the study of targets identified by modern genomic sequencing.
The MFED offers a more efficient approach to ligand generation for biological research and drug discovery.