Capillary electrophoresis affinity

Capillary affinity electrophoresis (CAE) or affinity capillary electrophoresis (ACE) — An electrophoretic separation technique (- electrophoresis), in which -> analytes are separated in a capillary, with the -> supporting (background) electrolyte containing substances capable of specific, often biospecific, interactions with the analytes. Ref [i] Riekkola ML, Jonsson jA, Smith RM (2004) Pure Appl Chem 76 443... 

In addition to open-tubular capillary zone electrophoresis, other modes of CE have been used for the separation of proteins and peptides capillary gel electrophoresis, capillary isoelectric focusing, and capillary affinity electrophoresis (406-417). 

Capillary affinity electrophoresis (CAE) based on specific carbohydrafe-protein interaction, is a valuable method in carbohydrate characterization. A variety of carbohydrate-binding proteins (e.g., lectins) specific to certain oligosaccharide structures are available and have been exploited by different research groups to characterize glycoprotein pharmaceuticals [145,146],... 

Nakajima, K., Oda, Y., Kinoshita, M. andKakehi, K. Capillary affinity electrophoresis for the screening of post-translational modification of proteins with carbohydrates. J Proteome Res, 2, 81, 2003. 

Nakajima, K. et al. Screening method of carbohydrate-binding proteins in biological sources by capillary affinity electrophoresis and its apphcation to determination of Tulipa gesneriana agglutinin in tulip bulbs. Glycobiology, 14, 793, 2004. 

Amini, A., Pettersson, C., and Westerlund, D. (1997). Enantioresolution of disopyramide by capillary affinity electrokinetic chromatography with human alphal-acid glycoprotein (AGP) as chiral selector applying a partial filling technique. Electrophoresis 18, 950—957. 

Capillary zone electrophoresis (CZE) is the most common electrophoretic separation technique due to its simplicity of operation and its flexibility. It is the standard mode for drug analysis, identification of impurities, and pharmacokinetic studies. Other separation modes, such as capillary isotachopho-resis (CITP), micellar electrokinetc chromatography (MEKC), capillary electrochromatography (CEC), capillary gel electrophoresis (CGE), capillary isoelectric focusing, and affinity capillary electrophoresis (ACE), have then-advantages in solving specific separation problems, since the separation mechanism of each mode is different. 

T Arai, M Ichinose, H Kuroda, N Nimura, T Kinoshita. Chiral separation by capillary affinity zone electrophoresis using an albumin-containing support electrolyte. Anal Biochem 217 7-11, 1994. 

T Arai, N Nimura, T Kinoshita. Investigation of enantioselective ofloxacin-albumin binding and displacement interactions using capillary affinity zone electrophoresis. Biomed Chromatogr 9 68—74, 1995. 

X Wu, RJ Linhardt. Capillary affinity chromatography and affinity capillary electrophoresis of heparin-binding proteins. Electrophoresis 19 2650-2653, 1998. 

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. 

Among the electrophoretic methods of chiral resolution, various forms of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary isotachophoresis (CIF), capillary gel electrophoresis (CGE), capillary isoelectric focusing (CIEF), affinity capillary electrophoresis (ACE), and separation on microchips have been used. However, in contrast to others, the CZE model has been used frequently for this purpose [44]. On the other hand, drawbacks associated with the electrophoretic technique due to lack of development of modem chiral phases have limited the application of these methods. Moreover, the electrophoretic techniques cannot be used at the preparative scale, which represents an urgent need of chiral separation science. 

Mangru, S.D., Harrison, D.J., Chemiluminescence detection in integrated postseparation reactors for microchip-based capillary electrophoresis and affinity electrophoresis. Electrophoresis 1998, 19(13), 2301-2307. 

The aim of this chapter is to cover the theoretical and practical aspects of capillary gel electrophoresis. It also provides an overview of the key application areas of nucleic acid, protein, and complex carbohydrate analysis, affinity-based methodologies, as well as related microseparation methods such as ultra-thin-layer gel electrophoresis and electric field-mediated separations on microchips. It also gives the reader a better understanding of how to utilize this technology, and determine which actual method will provide appropriate technical solutions to problems that may have be perceived as more fundamental. Micropreparative aspects and applications are discussed in Chapter 12. 

The next four chapters discuss the basic principles underlying operation and method development of the most common electrodriven analytical techniques CE, capillary isoelectric focusing (cIEF), capillary gel electrophoresis (CGE), and affinity capillary electrophoresis (ACE). Weinberger presents a comprehensive approach for method development in CE with an emphasis on small-molecule applications. This is followed by Kilar s chapter describing the principles of and method development in cIEF, as well as recent innovations... 

Guttman A, Cooke N (1991a) Capillary gel affinity electrophoresis of DNA fragments. Anal Chem 63 2038-2042. 

Ljunberg H and Nilsson S. Protein-based capillary affinity gel electrophoresis for chiral separation of 3-adrenergic blockers. 7. Liq. Chromatogr. 1995 18 3685. 

---