Peptide analytical chemistry

Hakansson, K., Emmett, M.R., Hendrickson, C.L. and Marshall, A.G. (2001) High-sensitivity electron capture dissociation tandem FTICR mass spectrometry of microelectrosprayed peptides, Analytical Chemistry 73,... 

Figure 7 CZE separation of hGH and its derivatives. CZE performed in FS capiiiary (i.d. 50 pm, totai iength 105cm, effective iength (to detector) 81.5cm BGE lOmmoii" Tricine, 5.8mmoii morphoiine, 20mmoii NaCi, pH8.0 eiectric fieid intensity 300Vcm current 20pA, temperature 24°C. 1, hGH 2, (desamido-149)- and (desamido-152)-hGH 3, (didesamido-149-152)-hGH. Mesityl oxide, electroosmotic fiow marker. (Reprinted with permission from Grossman PD, Coiburn JC, Lauer HH, et al. (1989) Application of free-solution capillary electrophoresis to the anaiyticai scaie separation of proteins and peptides. Analytical Chemistry 61(11) 1186-1194 American Chemical Society.)...

Figure 9.10 Three-dimensional representation of the data volume of a tryptic digest of ovalbumin. Series of planar slices through the data volume produce stacks of disks in order to show peaks. Reprinted from Analytical Chemistry, 67, A. W. Moore Jr and J. W. Jorgenson, Comprehensive three-dimensional separation of peptides using size exclusion chromatogra-phy/reversed phase liquid chromatography/optically gated capillary zone electrophoresis, pp. 3456-3463, copyright 1995, with permission from the American Chemical Society.

A Brief Review of the QSAR Technique. Most of the 2D QSAR methods employ graph theoretic indices to characterize molecular structures, which have been extensively studied by Radic, Kier, and Hall [see 23]. Although these structural indices represent different aspects of the molecular structures, their physicochemical meaning is unclear. The successful applications of these topological indices combined with MLR analysis have been summarized recently. Similarly, the ADAPT system employs topological indices as well as other structural parameters (e.g., steric and quantum mechanical parameters) coupled with MLR method for QSAR analysis [24]. It has been extensively applied to QSAR/QSPR studies in analytical chemistry, toxicity analysis, and other biological activity prediction. On the other hand, parameters derived from various experiments through chemometric methods have also been used in the study of peptide QSAR, where partial least-squares (PLS) analysis has been employed [25]. 

HPLC is frequently employed in the analysis of amino acids, peptides, proteins, nucleic acids, and nucleotides. HPLC is also often used to analyze for drugs in biological samples (see Workplace Scene 16.2). Due to the complex nature of the molecules to be analyzed, these techniques tend to be more complex than HPLC applications in other areas of analytical chemistry. For example, separation of nucleotides or amino acids is more difficult than testing for caffeine in beverages, even though the same instrument and same general methods would be employed. A variety of columns and mobile phases are regularly employed. 

G. J. Wegner, H. J. Lee, and R. M. Com, "Characterization and Optimization of Peptide Arrays for the Study of Epitopeantihody Interactions Using Surface Plasmon Resonance imaging," Analytical Chemistry 74, 5161-5168 (2002). 

Hydrazine sulfate is used as a reducing agent in analytical chemistry for gravimetric measurement of nickel, cobalt, and other metals, and in peptide analysis in the separation of polonium from tellurium as an antioxidant in... 

Yin, H., Killeen, K., Brennen, R., Sobek, D., Werlich, M., and van de Goor, T., Microfluidic chip for peptide analysis with an integrated HRLC column, sample enrichment column, and nanoelectrospray tip, Analytical Chemistry 11(2), 527-533, 2005. 

Moore, R. E., Licklider, L., Schumann, D., and Lee, T. D., A microscale electrospray interface incorporating a monolithic, poly(styrene-divinylbenzene) support for on-line liquid chromatography tandem mass spectrometry analysis of peptides and proteins. Analytical Chemistry 70(23), 4879-4884, 1998. 

Huang, X. A., Zhang, S., Schultz, G. A., and Henion, J., Surface-alkylated polystyrene monolithic columns for peptide analysis in capillary liquid chromatography-electrospray ionization mass spectrometry. Analytical Chemistry 74(10), 2336-2344, 2002. 

Martin, S. E., Shabanowitz, J., Hunt, D. R, and Marto, J. A., Subfemtomole MS and MS/MS peptide sequence analysis using nano-HPLC micro-ESI Eourier transform ion cyclotron resonance mass spectrometry, Analytical Chemistry 72(18), 4266 274, 2000. 

Timperman, A. T. and Aebersold, R., Peptide electroextraction for direct coupling of in-gel digests with capillary LC-MS/MS for protein identification and sequencing. Analytical Chemistry 72( 17), 4115-4121, 2000. 

With the advent of HPLC, a new tool was born that revolutionized peptide/protein chemistry as a whole as it allowed not only purification of biologically active peptides and proteins from complex mixtures of tissue or plant extracts/231 but also allowed purification of synthetic unprotected peptide mixtures analytically and preparatively.124-261 This was particularly significant in that purity could be assessed of peptide intermediates made by the classical solution-phase methodology that promoted characterization of all intermediates, as well as the purity of final products made by the solid-phase approach of MerrifieldJ27 ... 

Combinatorial chemistry has matured from a field where efforts initially focused on peptide-based research to become an indispensable research tool for molecular recognition, chemical-property optimization, and drug discovery. Originally used as a method to primarily generate large numbers of molecules, combinatorial chemistry has been significantly influenced and integrated with other important fields such as medicinal chemistry, analytical chemistry, synthetic chemistry, robotics, and computational chemistry. 

Fajer, P. G. (2000). Electron spin resonance spectroscopy labeling in proteins and peptides analysis. In Encyclopedia of Analytical Chemistry, (R. Meyers, ed.), pp. 5725-5761. Wiley, Chichester. 

Wilkins MR, Williams KL, Appel RD, Hochstrasser DF (1997) Proteome research new frontiers in functional genomics. Springer Verlag, Berlin Heidelberg Yates JR, Speicher S, Griffin PR, Hunkapiller T (1993) Peptide mass maps A highly informative approach to protein identification. Anal Biochem 214 397—408 Yates JR, Eng JK, McCormack AL, Schieltz D (1995) Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Analytical Chemistry 67 1426-1436. 

The enzymes used for this type of digestion in Analytical Chemistry are mainly hydrolytic enzymes, the catalytic effect of which is based on the insertion of water at a specific bond of the substrate. The hydrolytic enzymes used in analytical applications include lipases (which hydrolyse fats into long-chain fatty acids and glycerol) amylases (which hydrolyse starch and glycogen to maltose and to residual polysaccharides) and proteases (which attack the peptide bonds of proteins and peptides themselves). 

This is an undergraduate and introductory postgraduate textbook that gives information on amino acids and peptides, and is intended to be self-sufficient in all the organic and analytical chemistry fundamentals. It is aimed at students of chemistry, and allied areas. Suggestions for supplementary reading are provided, so that topic areas that are not covered in depth in this book may be followed up by readers with particular study interests. 

X.H., Fernandes, J.R., Grant, D.H., Ozsoz, M., Beglieter, A., and Mowat, M. (1997) Mismatch sensitive hybridization detection by peptide nucleic acids immobilized on a quartz crystal microbalance. Analytical Chemistry, 69 (24), 5200-5202. 

K. Shimura, W. Zhi, H. Matsumoto and K. Kasai, Accuracy in the determination of isoelectric points of some proteins and a peptide hy capillary isoelectric focusing utility of synthetic peptides as isoelectric point markers. Analytical Chemistry, 72, 4747 4757 (2(X)0). 

A. R. Wheeler, H. Moon, C. J. Kim, J. A. Loo, and R. L. Garrell, Electrowetting-based microfluidics for analysis of peptides and proteins by matrix-assisted laser desorption/ionization mass spectrometry. Analytical Chemistry, vol. 76, no. 16, pp. 4833-4838, Aug. 2004. 

Blondelle, S. E., Perezpaya, E., Dooley, C. T., Pinilla, C. Houghten, R. A. (1995) Soluble Combinatorial Libraries of Organic, Peptidomimetic and Peptide Diversities. Trac-Trends in Analytical Chemistry 14, 83-92. 

Mass spectrometer There are several commercially available MALDI-MS apparatus. These have been recently reviewed m the section product review of Analytical Chemistry (497 A, 1995). The analysis of the relatively complex peptide profiles of nervous tissue requires a high resolution mass spectrometer, i.e., an apparatus equipped with a reflectron (or m the linear mode with a capacity for delayed ion extraction). 

Moseley, M. A., Deterding, L. J., Tomer, K. B., and Jorgenson, J. W., Determination of bioactive peptides using capillary zone electrophoresis/mass spectrometry. Analytical Chemistry 1991, 63, 109-114. 

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