By Chenchen Wang, Xueping Fang, Cheng S. Lee (auth.), Nicola Volpi, Francesca Maccari (eds.)
Capillary electrophoresis (CE) is a comparatively new separation method compatible for dealing with small quantities of pattern vitally important in bioanalytical study and in a number of medical, diagnostic, genetic, and forensic purposes. In Capillary Electrophoresis of Biomolecules: equipment and Protocols, professional researchers within the box offer key options to enquire CE concentrating on easy and complicated carbohydrates (polysaccharides), aminoacids, peptides and proteins, enzymes, and nucleic acids. in addition to functional approaches, reports discussing CE functions concerning bio(macro)molecules also are integrated. Written within the hugely profitable Methods in Molecular Biology™ sequence layout, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, without problems reproducible laboratory protocols, and key tips about troubleshooting and averting identified pitfalls.
Authoritative and functional, Capillary Electrophoresis of Biomolecules: equipment and Protocols presents the reader with the newest holiday throughs and enhancements in CE and CE concepts utilized to numerous sessions of bio(macro)molecules.
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Additional resources for Capillary Electrophoresis of Biomolecules: Methods and Protocols
Humana Press, New York, pp 23–52 3. Evangelista RA, Liu MS, Chen FTA (1995) Characterization of 9-aminopyrene-1,4,6trisulfonate-derivatized sugars by capillary electrophoresis with laser-induced fluorescence detection. Anal Chem 67:2239–2245 4. Lv Z, Sun Y, Wang Y, Jiang T, Yu G (2005) Ultrasensitive capillary electrophoresis of oligoguluronates with laser-induced fluorescence detection. Chromatographia 61:615–618 5. García Moreno MDV, Castro Mejías R, Natera Marín R, García Barroso C (2002) Analysis of sugar acids by capillary electrophoresis with indirect UV detection.
Kazarian et al. used a low-pH sample (formic acid) and high-pH electrolyte (borate) to improve the sensitivity of carbohydrates modified with novel fluorescein reagent (40–42). This strategy was chosen because the derivatization reaction occurs at low pH and can be directly injected into the capillary, thereby avoiding the dilution step that is normally required. A 515fold improvement in sensitivity was obtained (Fig. 8) and was later implemented in microchips. One of the advantages of stacking with a dynamic pH junction is that it is tolerant of salt and is therefore compatible with highly saline solutions.
They applied their method to analysis of a proteinase K digest of thyroid-stimulating hormone (TSH). This glycoprotein contains several sulfonylated N-glycans including the 4-sulfonylated-LDn type that contains sulfonylated GalNAc in the antennae. The CE/MS data, acquired using negative-ion electrospray, showed abundant ions, demonstrating effective enrichment, from sulfonylated glycopeptides. The authors also showed that the glycopeptides could be analyzed using positive-ion electrospray MS by including a basic peptide of sequence KKK in the sheath liquid.