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Afi-Chip: An Equipment-Free, Low-Cost, and Universal Binding Ligand Affinity Evaluation Platform
Yanling Song1, Yuanzhi Shi1, Xingrui Li1
1MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, the 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, Department of Chemical Engineering, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University , Xiamen 361005, China.
Afi-Chip is a novel, equipment-free device for rapid and low-cost aptamer affinity evaluation. This portable platform simplifies molecular interaction studies, making aptamer screening more accessible.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Biosensing
Background:
- Characterizing binding affinity, specifically the dissociation constant (Kd), is crucial for aptamer screening and molecular interaction evaluation.
- Conventional methods for Kd determination often necessitate specialized equipment and extensive processing times, limiting their accessibility.
- There is a need for rapid, cost-effective, and equipment-free platforms for evaluating aptamer affinity.
Purpose of the Study:
- To introduce Afi-Chip, a portable, equipment-free platform for rapid and low-cost aptamer affinity evaluation.
- To demonstrate the versatility of Afi-Chip in characterizing aptamer binding affinities against diverse targets.
- To validate the performance of Afi-Chip by comparing its results with established methods.
Main Methods:
- Afi-Chip utilizes an enzyme-catalyzed reaction (H2O2 decomposition) to generate gas, driving an ink bar movement for a distance readout.
- The platform translates molecular recognition events into a measurable distance signal, enabling quantitative analysis.
- The system integrates reagent mixing and on-chip quantitative readout for streamlined analysis.
Main Results:
- Afi-Chip successfully monitored aptamer evolution and characterized binding affinities for targets including glucose, EpCAM protein, and SW620 tumor cells.
- The platform demonstrated generality by rapidly characterizing the affinity between anti-HCG and HCG.
- Kd values obtained using Afi-Chip were comparable to those determined by conventional methods and sophisticated instruments like flow cytometry.
Conclusions:
- Afi-Chip provides a novel, equipment-free approach for investigating molecular interactions.
- The platform offers a rapid, low-cost, and universal solution for aptamer affinity evaluation.
- Afi-Chip has potential applications in aptamer identification, ligand selection monitoring, and drug screening.
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