Bottom-up proteomics allows for rapid characterization of proteins by peptide mass fingerprinting, exploiting the specificity of enzymatic protein digestion and mass spectrometry to determine primary protein sequence. Conventional enzymatic digestion protocols are limited by the requirement of long incubation times, significant sample handling, and difficulties arising from enzymatic autolysis.  However, recent studies have shown that microwave dielectric heating can greatly accelerate the rate of the process, reducing digestion times from hours to minutes. The dielectric heating effect induced by microwave radiation allows localized heat generation based on the inherent dielectric loss of the materials, and thus it is well-suited to rapid modulation of digestion conditions within in-line flow-through reactors.<br/>Here, we report on the capabilities of a novel microstrip heating cavity specifically designed for electrospray ionization mass spectrometry.  Conventional microwave heating systems do not necessarily take full advantages of the field focusing capabilities of microwave radiation or are suitable to handle small volumes of sample (μL). The novel microwave resonator described here creates a highly-focused microwave electric field (2.45 GHz) within a microliter volume, allowing precise and modulated control of the proteolytic digestion.  Moreover, the approach permits microwave acceleration of digestions within the nano electrospray tip emitter itself, permitting the solution to be then sprayed directly without further sample handling.  When coupled in-line with a flowing solution source and mass spectrometer, the new system allows the digestion and complete characterization and elucidation of a wide range of proteins peptides greatly reducing sample digestion times over conventional protocols.  The current capabilities of the system for applications in bottom-up proteomics and rapidly-modulated digestion of biopolymers in mass spectrometry will be examined.<br/>

Novel system for rapidly-modulated microwave-assisted enzymatic proteolysis

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.