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Auto-affitech: an automated ligand binding affinity evaluation platform using digital microfluidics with a

Jingjing Guo1, Li Lin1, Kaifeng Zhao1

  • 1MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, Key Laboratory of Chemical Biology of Fujian Province, State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China. ljzhou@xmu.edu.cn cyyang@xmu.edu.cn.

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|March 25, 2020
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Summary
This summary is machine-generated.

This study introduces Auto-affitech, an automated platform using digital microfluidics to rapidly evaluate ligand binding affinity (Kd). It overcomes traditional method limitations, enabling faster, more accurate molecular recognition analysis.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Microfluidics

Background:

  • Dissociation constant (Kd) is vital for molecular recognition, but traditional methods are slow and error-prone.
  • Conventional Kd determination involves multi-step incubation, washing, and detection, leading to delays and potential errors.
  • Sensitive molecules can degrade, and manual operations increase the risk of personal errors during Kd analysis.

Purpose of the Study:

  • To develop an automated platform for rapid ligand binding affinity evaluation.
  • To overcome limitations of conventional methods for dissociation constant (Kd) determination.
  • To demonstrate a novel digital microfluidics (DMF) approach for efficient molecular binding analysis.

Main Methods:

  • Developed Auto-affitech, an automated ligand binding affinity evaluation platform utilizing digital microfluidics (DMF).
  • Implemented a novel bidirectional magnetic bead separation strategy within DMF for improved bead retention.
  • Programmed microliter-level droplets for rapid incubation and separation of target-beads and binding ligands.

Main Results:

  • Achieved high bead retention efficiency (89.57% ± 0.05%) and washing efficiency (99.59% ± 0.17%) using the bidirectional separation method.
  • Successfully determined the dissociation constant (Kd) for both aptamer-protein and antigen-antibody systems.
  • Demonstrated the platform's capability and universality across various receptor-ligand interactions.

Conclusions:

  • Auto-affitech provides an automated, efficient solution for dissociation constant (Kd) evaluation.
  • The platform minimizes manual labor, personal errors, sample consumption, and analysis time.
  • Auto-affitech shows significant potential for high-throughput screening of ligands and molecular recognition studies.