MS Analysis of Proteins

The ongoing development of top-down protein MS/MS capabilities (MS/MS) should prove quite valuable to researchers looking to identify and characterize proteins fractionated by 2DLC separations. Such methods are currently limited by restrictions on the maximum size of proteins analyzed, as well as analysis time-requirements that limit coupling of these methods with online LC analysis. Investigators from labs, such as Kelleher, McLafferty, Hunt (Coon et al., 2005 Meng et al., 2005 Han et al., 2006), and others are rapidly addressing these issues, and their methods will likely be adopted by many other researchers over the next few years. 

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Ivanov, A.R., Zang, L., Karger, B. L. (2003). Low-attomole electrospray ionization MS and MS/ MS analysis of protein tryptic digests using 20 pm-i.d. polystyrene-divinylbenzene monolithic capillary columns. Anal. Chem. 75, 5306-5316. 

Metabolic labeling is an in situ method which employs or isotope-encoded ( C or H) amino acids. Organisms are grown in controlled media under defined conditions in order to ensure translation of mRNA entities to uniformly isotopically encoded proteins. Consequently, the approach is generally restricted to cell culture systems and is not applicable to quantitative MS analysis of proteins derived from... 

Mane, C., Sommerer, N., Yalcin, T, Cheynier, V., Cole, R. B., Fulcrand, H. (2007a). Assessment of the molecular weight distribution of tannin fractions through MALDl-TOF MS analysis of protein-tannin complexes. Anal. Chem., 79, 2239-2248. 

LC/MS analysis of proteins and peptides is an important part of drug discovery process, as illustrated in this chapter. The combination of various HPLC techniques and advanced MS methods provides unique analytical capabilities of structural identifications for therapeutic proteins and target proteins. The continuous evolution of proteomics research provides both an opportunity and a challenge for further developments in separation techniques and MS characterization methods. It is expected that these analytical techniques will continue to play important roles in drug discovery in the future. 

Y. Zhao, W. Zhang, M.A. White, Y. Zhao, Capillary CE-MS analysis of proteins from affinity-purified plasma membrane. Anal. Chem., 75 (2003) 3751. 

K.C. Hansen, G. Schmitt-Ulms, R.J. ChaMey, J. Hirsch, M.A. Baldwin, A.L. Burlingame, MS analysis of protein mixtures at low levels using cleavable C-ICAT and multidimensional LC, Mol Cell Proteomics, 2 (2003) 299. 

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