By Nguyet Thuy Tran, Myriam Taverna
This e-book offers a complete survey of modern advancements and purposes of excessive functionality capillary electrophoresis within the box of protein and peptide research with a special specialise in the research of intact proteins. With useful element, the contents conceal various modes of capillary electrophoresis (CE) worthy for protein and peptide research, CZE, CIEF, ACE, CGE, and kinds of program reminiscent of the standard keep watch over of healing proteins and monoclonal antibodies, scientific analyses of chemokines in tissues, qualitative and quantitative research of vaccine proteins, and backbone of binding constants in complexes regarding peptides or proteins. Written for the hugely profitable Methods in Molecular Biology sequence, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and tips about troubleshooting and heading off identified pitfalls.
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Extra info for Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols
7 times). 18. Isopropanol is not required for the ionization of the proteins in sheathless CE-MS but it provides slightly better signal intensities for the particular proteins analyzed. References 1. Patrick JS, Lagu AL (2001) Review applications of capillary electrophoresis to the analysis of biotechnology-derived therapeutic proteins. Electrophoresis 22:4179–4196 2. Haselberg R, de Jong GJ, Somsen GW (2007) Capillary electrophoresis-mass spectrometry for the analysis of intact proteins. J Chromatogr A 1159:81–109 3.
7. Observe change in the fluorescent signal by injecting a fluorophore solution such as fluorescein and rhodamine B with a microsyringe to confirm whether alignment of the detection system is correct or not. 18 3 Takashi Kaneta Methods 1. 1 M NaOH for 5 min, deionized water for 5 min, and the running buffer for 5 min, sequentially. 2. Condition the reaction capillary (large bore capillary) by rinsing with water and the running buffer sequentially. 3. Fill the separation capillary and the reaction capillary with the running buffer and the derivatization solution, respectively (see Note 14).
Compared to LC, CE provides a higher separation efficiency and more important a different selectivity (charge/size-ratio). Thus, it is often possible to separate protein isoforms like glycoforms. Moreover, proteins are easily charged in acidic background electrolyte (BGE) and therefore ideal for CE separation. For CE-MS coupling, electrospray ionization (ESI) is applicable due to the ionic character of the analyzed proteins. ), Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols, Methods in Molecular Biology, vol.