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A novel poly(deep eutectic solvent)-based magnetic silica composite for solid-phase extraction of trypsin
Kaijia Xu1, Yuzhi Wang1, Yixue Li1
1State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, PR China.
Analytica Chimica Acta
|November 9, 2016
Summary
Novel magnetic microspheres functionalized with poly(deep eutectic solvent) were developed for efficient trypsin extraction. This method offers high capacity, selectivity, and preserves enzyme activity, showing promise for biomolecule separation.
Area of Science:
- Materials Science
- Biochemistry
- Analytical Chemistry
Background:
- Magnetic microspheres offer advantages in separation due to ease of manipulation.
- Deep eutectic solvents (DES) are emerging as versatile media in various chemical applications.
- Trypsin is a crucial enzyme in biological research and industrial processes.
Purpose of the Study:
- To synthesize and characterize novel poly(deep eutectic solvent) grafted silica-coated magnetic microspheres (Fe3O4@SiO2-MPS@PDES).
- To investigate the application of these microspheres for magnetic solid-phase extraction (MSPE) of trypsin.
- To evaluate the extraction capacity, selectivity, and reusability of the developed material for trypsin purification.
Main Methods:
- Synthesis of Fe3O4@SiO2-MPS@PDES via polymerization of choline chloride-itaconic acid.
- Characterization using VSM, FT-IR, XPS, TGA, and TEM.
- Optimization of MSPE conditions (pH, ionic strength, temperature, time) for trypsin extraction.
- UV-Vis spectrophotometry for trypsin quantification.
- Activity assays to assess retained trypsin function.
Main Results:
- The Fe3O4@SiO2-MPS@PDES microspheres exhibited a high extraction capacity for trypsin, reaching up to 287.5 mg/g under optimal conditions.
- The developed material demonstrated superior extraction capacity and selectivity for trypsin compared to unmodified microspheres.
- The microspheres were recyclable for at least six cycles with minimal loss of extraction capacity, retaining 233 mg/g after eight cycles.
- The activity of the extracted trypsin was well preserved, and successful purification from crude bovine pancreas extract was achieved.
Conclusions:
- Novel Fe3O4@SiO2-MPS@PDES magnetic microspheres are effective for the MSPE of trypsin.
- The method provides high efficiency, selectivity, and reusability, with preserved enzyme activity.
- This approach holds significant potential for the separation and purification of biomolecules.
