Mass Spectrometry Analysis of Liquid Chromatography Fractions using Ettan LC-MS Systems

GUIDE TO LC–MS.
Summary Mass spectrometry provides a faster of fractions. The procedure described involves initial protein purification on ÄKTA™ purifier 10, automated and rapid desalting of fractions on Ettan microLC, and MS analysis by electrospray ionization time-of-flight mass spectrometry (ESI-ToF MS) using Ettan ESI-ToF. Ettan LC–MS provides high-quality mass spectra of a wide range of high molecular weight proteins including bovine serum albumin, cytochrome C, transferrin, ovalbumin, myoglobin, a monoclonal antibody and two -lactoglobulin variants.
 
Introduction Mass spectrometry offers an excellent means of monitoring protein purification and performing confirmation analysis of protein fractions collected from chromatographic purification. However, chromatography techniques normally used rely on the ion exchange, size exclusion or hydrophobic interaction properties of the proteins. With these techniques the mobile phase is ionic and at about neutral pH, (i.e., conditions that are poorly or not at all compatible with electrospray ionization mass spectrometry (ESI MS, Figure 1). We show results of an efficient technique for automated desalting in combination with mass analysis using Ettan ESI-ToF. The mass spectrometer is suited to mass analysis over a wide mass range. It is equipped with dual-ionizationand more accurate tool for analysing liquid chromatography fractions during protein purification than traditionally used techniques (e.g., electrophoresis). Liquid chromatography–mass spectrometry (LC–MS) using Ettan™ LC–MS offers an excellent means of monitoring protein purification by determining molecular weight, identity and purity,

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