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Poster Presentations
Day 4, June 25（Wed.）
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-AM-26
Development of a Miniature Ultrasonic Device for Rapid Enzymatic Digestion of Protein
(NCKU Chem)
oPo-Yu Chou, Szu-Hsueh Lai
Bottom-up proteomics is a widely used approach for protein identification and characterization of amino acid sequences and post-translational modifications (PTMs). This method involves enzymatic digestion of proteins into peptides prior to mass spectrometry analysis. The resulting peptides are then separated using one or more dimensions of liquid chromatography coupled with mass spectrometry. Peptide identification is performed by comparing observed peptide masses and tandem mass spectra with predicted data from sequence databases or spectral libraries. Multiple peptide identifications are subsequently assembled to infer protein identities. Nevertheless, conventional protein digestion is a time-intensive process, often requiring several hours to complete. Over the past two decades, various acceleration techniques have been developed to improve efficiency, such as microwave- and ultrasound-assisted methods. However, sample preparation remains laborious due to extensive pre-treatment steps for protein denaturation. To address this, we developed a cost-effective ultrasonic nebulization device for rapid protein digestion. This method digested myoglobin in just 2 minutes without prior denaturation, achieving 100% sequence coverage with low-enzyme-activity trypsin (<2,000 U) as confirmed by MALDI-TOF MS. In contrast, other methods typically require sequence-grade enzymes (>20,000 U). Our approach not only accelerates digestion and enhances efficiency but also significantly reduces enzyme costs.