By Charles A. Lucy, Nicole E. Baryla (auth.), Mark A. Strege, Avinash L. Lagu (eds.)
Capillary electrophoresis (CE) is likely one of the most desirable equipment for the fast research of pharmaceutical proteins and peptides, and therefore now much more very important with the sequencing of the human genome and the next introduction of proteomics. In Capillary Electrophoresis of Proteins and Peptides, prime educational scientists and professional laboratorians operating within the pharmaceutical describe their most sensible CE suggestions in step by step aspect for simple replica. The authors current state of the art innovations for either capillary coatings and analytical detection through laser-induced fluorescence, for the advance and commercialization of biopharmaceuticals, and for affinity capillary electrophoresis within the evaluate of protein binding, together with using protein cost ladders. extra chapters talk about CE and capillary isoelectric focusing, mixed with electrospray mass spectrometry detection, for appearing proteomic experiences. parts of certain curiosity lined comprise proteomic purposes, in addition to purposes within the box of protein-ligand binding. The protocols keep on with the winning tools in Molecular Biology™ sequence structure, each one providing step by step laboratory directions, an creation outlining the main at the back of the strategy, lists of kit and reagents, and pointers on troubleshooting and fending off identified pitfalls.
complete and cutting-edge, Capillary Electrophoresis of Proteins and Peptides will end up necessary to either new and skilled investigators engaging in major protein and peptide experiences in biotechnology, drug discovery, and pharmaceutical examine and development.
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Extra info for Capillary Electrophoresis of Proteins and Peptides
The experiment involves micellar electrokinetic capillary chromatography (MEKC) using a borate buffer with sodium dodecyl sulfate (SDS) and tetrahydrofuran (THF) additives. MEKC is a technique that involves the use of charged micelles to separate charged and uncharged molecules by means of a pseudomicellar phase that is created through hydrophobic interactions between solute molecules and detergent. The CE/MEKC method (17) was used to identify 18 CBQCAderivatized amino acids (see Fig. 4). Seventeen of them were quantitated at attomolar concentrations (see Table 1).
The separation conditions of enantiomers using natural β-cyclodextrins added to a phosphate buffer containing a small amount of organic modifier were similar to conditions developed by Ward et al. (18). In order to get a faster enantiomer separation, the method was modified by using a shorter capillary, buffer pH was lowered, and the β-cyclodextrin concentration was raised. The experiment was carried out in darkness to prevent interference from ambient light, thereby increasing the exposure time and subsequently lowering the limit of detection.
Alberta, Canada, Chs. 1 and 6. 2. , and Moring, S. (1991) Fluorescence detection in capillary electrophoresis: evaluation of derivatizing reagents and techniques. Anal. Chem. 63, 417–422. 3. 3 Wu, S. and Dovichi, N. J. (1989) High-sensitivity fluorescence detector for fluoresceinisothiocyanate derivatives of amino acids separated by capillary electrophoresis. J. Chromatogr. 480, 141–155. 4. 4 Dovichi, N. , Martin, J. , Jett, J. , and Keller R. A. (1984) Laserinduced fluorescence of flowing samples as an approach to single-molecule detection in liquids.
Capillary Electrophoresis of Proteins and Peptides by Charles A. Lucy, Nicole E. Baryla (auth.), Mark A. Strege, Avinash L. Lagu (eds.)