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Updated: Jul 10, 2026
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Crystallization and Structural Determination of an Enzyme:Substrate Complex by Serial Crystallography in a Versatile Microfluidic Chip
Published on: March 20, 2021
Crystalline COFs Assembled in an Enzyme-Compatible Milieu for Cascade Biosensing
Dongxue Feng1, Yuxin Zhou1, Yao Tong1
1Key Laboratory for Ultrafine Materials of Ministry of Education, School of Materials Science and Engineering, East China University of Science and Technology, Shanghai 200237, China.
ACS Applied Materials & Interfaces
|July 9, 2026
Summary
This study introduces a novel method for creating enzyme-encapsulating covalent organic frameworks (COFs) in water. This new COF platform enables sensitive detection of chronic kidney disease biomarkers, offering a promising tool for early diagnosis.
Area of Science:
- Materials Science
- Nanotechnology
- Biochemistry
Background:
- Covalent organic frameworks (COFs) assembly in enzyme-compatible media is challenging.
- Enzyme encapsulation within COFs is crucial for developing advanced biosensors.
- Developing additive-free, aqueous synthesis methods for COFs is highly desirable.
Purpose of the Study:
- To develop an electron donor-acceptor (EDA)-complex-assisted strategy for aqueous, additive-free synthesis of β-ketoenamine COFs.
- To achieve simultaneous in situ enzyme encapsulation within the COFs.
- To create a spatially coupled nanoreactor for detecting chronic kidney disease (CKD) biomarkers.
Main Methods:
- Utilized an EDA complex formed by 2,2'-bipyridine-5,5'-diamine (Bpy) and 1,3,5-triformylphloroglucinol (Tp) for COF synthesis.
- Employed additive-free, aqueous ambient conditions for COF assembly and enzyme encapsulation.
- Designed a nanoreactor by encapsulating creatinine deiminase (CD) and installing a biomimetic laccase (Bpy-Cu).
Main Results:
- Achieved highly crystalline TpBpy COFs with enhanced error correction via EDA complex prearrangement.
- Successfully encapsulated enzymes within the COF structure, utilizing Bpy units for biomimetic enzymatic center creation.
- Developed a cascade amplification system for specific creatinine detection, achieving a low detection limit of approximately 7.7 μM.
- Demonstrated the platform's potential for assessing kidney function and monitoring CKD.
Conclusions:
- The EDA-complex-assisted strategy enables efficient, additive-free synthesis of enzyme-loaded COFs in aqueous media.
- The developed COF-based nanoreactor provides a sensitive platform for CKD biomarker detection.
- This modular and generalizable approach holds promise for the early diagnosis of various diseases by altering encapsulated enzymes and biomimetic sites.
