Download Analytical and Preparative Separation Methods of by Hassan Y. Aboul-Enein PDF

By Hassan Y. Aboul-Enein

Reviews updated study advancements on purifying and isolation huge natural molecules. The textual content offers info on high-performance liquid chromatography and capillary electrophoresis (CE) as instruments for examining biomacromolecules and constructing new biochemical and medicinal compounds. It applies biochemical separation know-how to the examine of macromolecules similar to proteins, polysaccharides, nucleic acids and extra.

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Consequently, from this region of maximal difference of charges the suitable pH of BGE should be chosen. For the given example of diglycine and triglycine it is evident that the suitable pH for their separation is either at low acid region (pH < 3) or at alkaline pH (pH > 9), since in these regions the differences of specific and corrected specific charges are maximal. 5, was chosen as BGE for the separation of these two peptides, which are often used as test mixture in our laboratory. The record of CZE separation of these two peptides and the electroosmotic flow marker, phenol, is given in Fig.

Recombinant Proteins and MS 37 Bischoff R, Roecklin D, Roitsch C. Electrophoresis 1992;13:214-219. Bouchon B. Van Dorsselaer A, Roitsch C. Biol Mass Spectrom 1993;22:358-360. Bouchon B, Klein M, Bischoff R, Van Dorsselaer A, Roitsch C. Anal Biochem 1997;246:52-61. Bouchon B, Jaquinod M, Klarskov K, Trottein F, Klein M, Van Dorsselaer A, Bischoff R, Roitsch C. J Chromatogr 1994;662:279-290. Wilm M, Mann M. Anal Chern 1996;68:1-8. Falick AM, Hines WM, Medzihradsky KF, Baldwin MA, Gibson BW. J Am Soc Mass Spectrom 1993;4:882-893.

The purified peptide ( < I pg) was collected for further analysis. Tryptic peptides are generally characterized during ESMS analysis by the presence of two ionization positions (The N terminus and the basic C-terminal residue, except for the C-terminal peptide of any digest). In the case of rOspA, however, the N-terminal peptide presented only one ionization site due to Nterminal blockage. The most abundant peak in this spectrum (Fig. 4 Da), indicating that one fatty acid was unsaturated. In addition, several minor components were detectable.

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