High performance capillary electrophoresis peptides

High performance capillary electrophoresis was introduced originally as an analytical tool. Now that instruments are equipped with automated fraction collection, however, capillary electrophoresis can be used for micropreparative collection of individual peaks separated from a mixture. Using the fraction collection feature, nanomolar amounts of solute such as proteins, peptides, oligonucleotides can be collected in amounts sufficient for microsequencing. An intersample washing procedure and use of well-formed capillaries aid in the prevention of artifacts.44... 

Boss, H. J., Rohde, M. F., and Rush, R. S., Multiple sequential fraction collection of peptides and glycopeptides by high-performance capillary electrophoresis, Anal. Biochem., 230, 123, 1995. 

Satow, T., Machida, A., Funakushi, K., and Palmieri, R., Effects of the sample matrix on the separation of peptides by high performance capillary electrophoresis, HRC CC, 14, 276, 1991. 

Finally, when RPC methods are used in preparative studies with peptides, the opportunity routinely exists for subsequent analysis of the recovered fractions by a variety of analytical methods including high-speed RP-HPLC, HP-IEX, HP-HILIC, or HP-IMAC, zonal or micellar electrokinetic high-performance capillary electrophoresis (HP-CZE and MECK-CZE), capillary electrochromatography (CEC), or capillary isotachophoresis. The combination of the RPC information, drawn from the In k versus i > plots, with the data derived from on-line spectroscopic detection thus readily provides a comprehensive opportunity to assess the purity of an isolated peptide, many of the physicochemical features of the interaction, as well as a means to optimize the resolution in the RPC separation. 

Boutin, J. A., Hennig, P., Lambert, P. H., et al. (1996) Combinatorial peptide libraries Robotic synthesis and analysis by nuclear magnetic resonance, mass spectrometry, tandem mass spectrometry, and high-performance capillary electrophoresis techniques. Anal. Biochem. 234, 126-141. 

B. Volland, J.P f auchbre, J.L. Combinatorial Peptide Libraries Robotic Synthesis and Analysis by Nuclear Magnetic Resonance, Mass Spectrometry, Tandem Mass Spectrometry, and High-performance Capillary Electrophoresis Techniques, AnflZ. Biochem. 234, 126-141 (1996). 

Rush R S, Derby P L, Strickland T W, et al. (1993). Peptide mapping and evaluation of glycopeptide microheterogeneity derived from endoproteinase digestion of erythropoietin by affinity high-performance capillary electrophoresis. Analyt. Chem. 65 1834-1842. 

Landers, J. R, Oda, R. R, Liebenow, J. A., and Spelsberg, T. C. Utility of high performance capillary electrophoresis for monitoring peptide homo- and hetero-dimer formation, J. Chrorrmtogr., 652,109-117,... 

Elsevier, 2005 Capillary Electrophoresis of Proteins and Peptides. M. A. Strege and A. L. I-agu, eds., Totowa, NJ Humana Press. 2004 Clinical and Forensic Applications of Capillary Electrophoresis. J. R. Petersen and A. A. Mohamad, eds., Totowa, NJ Humana Press. 2001 R. Weinberger. Practical Capillary Electrophoresis. 2nd ed New York Academic Press. 2000 High Performance Capillary Electrophoresis. M, G. Khaledi. ed., New brk Wiley. 

The application of high-performance capillary zone electrophoresis (HP-CZE) in its various selectivity modes has become a very valuable adjunct to HPLC for the analysis of peptides. For synthetic peptides, in particular, both HPLC and HP-CZE now form essential components of the analytical characterization of these molecules. Increasingly, zonal, micellar, or (biospecific) affinity-based HP-CZE procedures with open tubular capillary systems are adapted to allow resolution with extremely high separation efficiencies (e.g., >105 plates per meter) of synthetic or naturally occurring peptides as part of the determination of their structural, biophysical, or functional properties. Illustrative of these capabilities are the results shown in Figure 19 for the separation of several peptides with different charge and Stokes radius characteristics by HP-CZE. 

High performance liquid chromatography (HPLC) and capillary electrophoresis (CE) are two instrumental separation techniques that are applicable to the separation of proteins and peptides. The advantage of HPLC and CE techniques is that they afford the analyst the freedom to resolve a complex mixture by different routes employing different... 

C Miller, J Rivier. Peptide chemistry development of high performance liquid chromatography and capillary zone electrophoresis. Biopolymers (Pept Sci) 40, 265, 1996. 

Ridge, S., and Hettiarachchi, K. (1998). Peptide purity and counter ion determination of bradykinin by high-performance liquid chromatography and capillary electrophoresis. /. Chromatogr. A 817, 215-222. 

KM De Antonis, PR Brown. Analysis of derivatized peptides using high performance liquid chromatography and capillary electrophoresis. Adv Chromatogr 37 425-452, 1997. 

A. J. Tomlinson and S. Naylor, Enhanced performance membrane preconcentration-capillary electrophoresis-mass spectrometry (mPC-CE-MS) in conjunction with transient isotachophoresis for analysis of peptide mixtures,./. High Resolut. Chromatogr. 18 384-386(1995). 

The high sample demands and low-throughput of LC-MS methods have led to the creation of a capillary electrophoresis (CE) platform for ABPP [48]. Proteomes are labeled with a fluorescent probe, digested with trypsin, and enriched with antifluorophore antibody resins. Use of CE coupled with laser-induced fluorescence (LIF) detection to analyze the enriched peptides resulted in far superior resolution to ID SDS-PAGE, particularly for enzymes that share similar molecular masses. Sensitivity limits of 0.05-0.1 pmol/mg proteome, negligible sample requirements (—0.01—0.1 pg proteome), and the ability to perform rapid CE runs in parallel with 96-channel instruments, make CE-based ABPP a potentially powerful technique. One drawback is that the identities of the probe-labeled proteins are not immediately apparent, and correlated LC-MS experiments must be performed to assign protein identities to the peaks on the CE readout. 

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