Development of a 20 kpsi Enzymatic Digester for High Throughput Proteomic Analysis and its Application to Membrane Proteomics

摘要

A 20 kpsi enzymatic digester was developed and performance demonstrated using a 4-protein standard and hydrophobic membrane proteins. The digester was shown to be robust and compatible with nano-flow LC-MS/MS, enabling online enzymatic lysis of hydrophobic membrane proteins. For the S. oneidensis insoluble fraction, the sample digested for 20 min in the PCT reactor showed more protein identifications than the sample digested overnight. The ultra-high pressure PCT reactor facilitates proteomics research demanding high throughput technology, especially membrane protein analysis. Incorporation of protein digestion into a fully automated online LC-MS platform reduces sample handling errors and losses associated with offline processing methods.