An integrated microfluidic system with shear force control for an automatically modified mRNA display technique for...

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Biology

Background:

• Cancer research extensively explores KIF2C protein for therapeutic development.
• Traditional peptide probe screening is time-consuming and labor-intensive.
• Modified messenger RNA (mRNA) display is a powerful technique for identifying high-affinity ligands.

Purpose of the Study:

• To develop a low-cost, automated microfluidic system for rapid peptide probe screening.
• To optimize the modified mRNA display technique for high-specificity peptide selection.
• To identify novel peptide probes targeting the cancer-overexpressed KIF2C protein.

Main Methods:

• Development of an integrated microfluidic system (IMS) with micropumps, microvalves, micromixers, and thermoelectric modules.
• Automation of the modified mRNA display technique on a single microfluidic chip.
• Precise control of shear force during washing steps to eliminate off-target peptide binding.

Main Results:

• The IMS successfully automated modified mRNA display, reducing screening time from days to under one day.
• Precise shear force control (0.96–9.6 nN) effectively eliminated off-target peptides under physiological conditions.
• One identified peptide, OCTRD 14, showed high binding affinity and specificity for KIF2C, with a dissociation constant (Kd) of 2.4 μM, comparable to antibody-like binding.

Conclusions:

• The developed IMS offers a high-efficiency, automated platform for peptide probe screening.
• This technology significantly accelerates the discovery of targeted peptide therapeutics.
• OCTRD 14 represents a promising antibody-like candidate for KIF2C-targeted cancer therapies.