By Nguyet Thuy Tran, Myriam Taverna
This ebook presents a entire survey of contemporary advancements and purposes of excessive functionality capillary electrophoresis within the box of protein and peptide research with a special concentrate on the research of intact proteins. With sensible element, the contents hide varied modes of capillary electrophoresis (CE) priceless for protein and peptide research, CZE, CIEF, ACE, CGE, and kinds of program akin to the standard keep watch over of healing proteins and monoclonal antibodies, medical analyses of chemokines in tissues, qualitative and quantitative research of vaccine proteins, and resolution of binding constants in complexes concerning peptides or proteins. Written for the hugely winning Methods in Molecular Biology sequence, chapters contain introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and pointers on troubleshooting and warding off identified pitfalls.
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Extra resources for Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols
G. immunogenicity), thereby compromising their therapeutic properties. Quality control of biopharmaceuticals is an essential—but challenging—task, requiring analytical techniques allowing characterization of intact proteins and their variants. ), Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols, Methods in Molecular Biology, vol. 1007/978-1-4939-4014-1_3, © Springer Science+Business Media New York 2016 25 26 Elena Domínguez-Vega et al. proteins . CE is carried out in an open narrow tube—usually a fused-silica capillary—filled with a background electrolyte solution.
36 Elena Domínguez-Vega et al. 0 time (min) Fig. 4 (a) Schematic representation of the production of drug–protein conjugates. 1) BGE containing 5 % (v/v) of isopropanol. (c) Molecular mass and assigned conjugates obtained for peaks 1–6. Reprinted with permission from ref.  12. Monitor the BPE obtained in the range of 1000–1800 m/z. Figure 4b shows the BPE obtained for the erlotinib-LZM conjugates under these conditions. 13. Determine the mass of the observed peaks by deconvolution of the mass spectra.
1) Complex between thyroglobulin and Rh-Con A, (2) albumin, (3) fluorescein. Blue lines and green lines are obtained by 450-nm excitation and 532-nm excitation, respectively. Reproduced from  with permission from John Wiley and Sons 5. Employ molecular masses of 660 and 66 kDa for thyroglobulin and albumin, respectively, to calculate the concentrations of the protein solutions. 6. Prepare fresh samples when the peaks of proteins are distorted. 7. 9 × 10−5 M, using molecular mass of 102,000 for concanavalin A.