Electrospray ionization mass spectrometry recombinant proteins

Tang, Q., Harrata, A. K., Lee, C. S. (1997). Two-dimensional analysis of recombinant E. coli proteins using capillary isoelectric focusing electrospray ionization mass spectrometry. Anal. Chem. 69(16), 3177-3182. 

K.L.Johnson, T.D. Veenstra,J.M. Londowski, A.J.Tomlinson, R. Kumar, S. Naylor, On-line sample clean-up and chromatography coupled with electrospray ionization mass spectrometry to characterize the primary sequence and disulfide bond content of recombinant calcium binding proteins, Biomedical Chromatography 13 (1999)37-45. 

Coupling CE with electrospray ionization mass spectrometry (MS) can potentially be a very powerful tool for detecting and identifying product-related impurities in recombinant pharmaceuticals. Proteins can be detected in the femtomole range with this mode. Conceivably, the bioanalyst could perform a peptide map with CZE-MS and detect, identify, and sequence aberrant peptides derived from degradates. 

Troxler, H., et al. (1999). Electrospray Ionization Mass Spectrometry Analysis of the Ca -binding of Human Recombinant a-parvalbumin and Nine Mutant Proteins, Anal. Biochem. 268 64-71. 

Recombinant wild type hIL-ip, the K138C mutant and the K138C, R4A, L6A triple mutant (mutant 1) were isolated from the soluble fraction of E. coli lysates by ammonium sulfate fractionation and hydrophobic interaction chromatography. The purified proteins were characterized by SDS-PAGE, western blots, N-terminal sequence, size exclusion chromatography (SEC), isoelectric focusing (lEF), matrix assisted laser desorption ionization mass spectrometry (MALDI-MS), and electrospray mass spectrometry (ESMS). 

Fenn JB, Mann M, Meng CK Electrospray ionization for mass spectrometry of large biomolecules. Science (1989) 246 64-71. Patrick JS, Lagu AL Review applications of capillary electrophoresis to the analysis of biotechnology-derived therapeutic proteins. Electrophoresis (2001) 22 4179-4196. Sowell J, Salon J, Strekowski L, et al Covalent and noncovalent labeling schemes for near-infrared dyes in capillary electrophoresis protein applications. Methods Mol. Biol. (2004) 276 39-75. Moini M Capillary electrophoresis mass spectrometry and its application to the analysis ofbiological mixtures. Anal. Bio-anal. Chem. (2002) 373 466 180. Nemunaitis J, Holmlund JT, Kraynak M, et al. Phase I evaluation of ISIS 3521, an antisense oligodeoxynucleotide to protein kinase C-a, in patients with advanced cancer./. Clin. Oncol. (1999) 17 3586-3595. De Frutos M, Cifuentes A, Diez-Masa JC Differences in capillary electrophoresis profiles of urinary and recombinant erythropoietin. Electrophoresis (2003) 24 678-680. 

T Liu, XX Shao, R Zeng, QC Xia. Analysis of recombinant and modified proteins by capillary zone electrophoresis coupled with electrospray ionization tandem mass spectrometry. J Chromatogr A 855 695-707, 1999. 

Lupin proteins have been analyzed by a number of different techniques, including liquid chromatography (Duranti et al., 1995), differential sedimentation (Franco et al., 1997 Freitas et al., 2000), proteomic analysis by one- and two-dimensional gel electrophoresis (Duranti et al., 1992 Magni et al., 2005b) or liquid chromatogra-phy/electrospray ionization tandem mass spectrometry (Schwend et al., 2003 Wait et al., 2005 Magni et al., 2007), isoelectrofocusing (Quaresma et al., 2007), crystalb-zation (Biesiadka et al., 1999), and expression of recombinant protein (Scarafoni et al., 2001). 

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