Interface solution isoelectric focusing with in situ MALDI-TOF mass spectrometry

Abstract

This paper describes a simple and reusable microfluidic device combining solution IEF (sIEF) with MALDI-TOF MS for rapid proteomic and metabolic analysis of microliter samples. The device contains two glass plates with nanoliter microwell arrays, which can be assembled to form a fluidic path for sIEF separation, and reconfigured for dividing separated bands. One microliter samples can be loaded and separated by sIEF into static bands in 10∼30 min. After a slipping operation, the static IEF bands can be divided into nanoliter droplets in microwells without mobilization, and the device can be opened for in situ MALDI-TOF MS detection without loss of separation resolution. The performance of the device is characterized by separating and identifying intact proteins. The applicability in metabolic analysis is demonstrated by preliminary experiments on profiling small molecular metabolites in cerebrospinal fluid microdialysates from rat brain.

Keywords: MALDI; Microfluidics; Proteomics; SlipChip; Solution IEF.