Core-shell silica microsphere-based trypsin nanoreactor for low molecular-weight proteome analysis.

Area of Science:

• Materials Science
• Analytical Chemistry
• Biotechnology

Background:

• Core-shell mesoporous silica (CSMS) microspheres are crucial for bioapplications due to tunable mesopores.
• Efficient analysis of low molecular-weight (MW < 30 kDa) proteins remains a challenge in proteomics.

Purpose of the Study:

• To develop and analyze novel CSMS microspheres for size-exclusion separation and enzyme immobilization.
• To evaluate the performance of CSMS-based enzyme nanoreactors for low MW protein analysis.

Main Methods:

• Synthesis of CSMS microspheres via a two-phase process.
• Immobilization of trypsin onto CSMS using physical absorption or covalent binding.
• Analysis of protein digestion products using MALDI-TOF MS and capillary electrophoresis (CE).

Main Results:

• CSMS microspheres exhibited uniform size (1.3 μm), tunable mesopores (7.9 nm), high surface area (55.5 m²/g), and excellent water dispersibility.
• High trypsin loading capacity (11.8–6.1 mg/g) was achieved on CSMS.
• CSMS-based nanoreactors demonstrated size-dependent digestion, enhancing peptide-sequence coverage for smaller proteins.

Conclusions:

• Novel CSMS microspheres provide a promising platform for size-selective proteome analysis.
• The developed enzyme nanoreactors facilitate improved analysis of low MW proteins.
• This approach paves the way for advanced applications of mesoporous materials in proteomics.