High-performance liquid chromatography separation and intact mass analysis of detergent-solubilized integral membrane proteins

Journal article

We have developed a method for intact mass analysis of detergent- solubilized and purified integral membrane proteins using liquid chromatography-mass spectrometry (LC-MS) with methanol as the organic mobile phase. Membrane proteins and detergents are separated chromatographically during the isocratic stage of the gradient profile from a 150-mm C3 reversed-phase column. The mass accuracy is comparable to standard methods employed for soluble proteins; the sensitivity is 10-fold lower, requiring 0.2-5 μg of protein. The method is also compatible with our standard LC-MS method used for intact mass analysis of soluble proteins and may therefore be applied on a multiuser instrument or in a high-throughput environment. © 2010 Elsevier Inc. All rights reserved.

Authors: Berridge, G; Chalk, R; D'Avanzo, N; Dong, L; Doyle, D

• Journal: Analytical Biochemistry
• Volume: 410
• Issue: 2
• Pages: 272-280
• Publication date: 2011-03-15
• DOI: 10.1016/j.ab.2010.11.008
• EISSN: 1096-0309
• ISSN: 0003-2697
• Language: English
• Keywords: Intact mass analysis; Isocratic elution; LC-MS; HPLC; Electrospray ionization mass spectrometry; Detergent; Integral membrane protein; Methanol