Reversed Phase Monolithic Column Based Enzyme Reactor for Protein Analysis

We developed a reversed-phase monolithic column based enzyme reactor for protein analysis. The effects of buffer solution, porous structure of the column, digestion time and hydrophobicity of solid phase on the digestion performance of enzyme reactor were systematically investigated in terms of the number of identified unique peptides, sequence coverage and the level of missed cleavage as the evaluation indicators. The results demonstrated that the reversed-phase monolithic column based enzyme reactor had high digestion efficiency and might be further extended to the proteome analysis of complex samples.

Graphical abstractBase peak chromatogram for separation of 1 μg BSA digest by C12 monolithic column microreactor. This new enzyme reactor showed more base peaks and higher peak intensity in the peak chromatogram compared with C18 monolithic column and C18 packed column microreactors.Figure optionsDownload full-size imageDownload as PowerPoint slide