Capillary zone electrophoresis of proteins and neuroprotective peptides
|Kapilární zónová elektroforéza proteinů a neuroprotektivních peptidů
|Článek ve sborníku
|Third International and Twenty-Eighth European Peptide Symposium. Peptides 2004.
|Fakulta / Pracoviště MU
|Analytická chemie, separace
|capillary coating; neuroprotective peptides; capillary zone electrophoresis
|Biocompatibility of inner surface of capillaries in CE is critical for separation and detection of proteins and peptides. The problems come from adsorption of the peptides (proteins) on the surface because biomolecules are sensitive to changes in ionic strength, pH, and/or temperature of the electrolyte, etc. Therefore, modification of the fused silica capillary surface is often necessary. Procedures for capillary coating described in literature are usually complicated and time consuming. We optimized three types of relatively simple coating procedures suitable for protein and peptides separation: a/ polyethyleneimine (PEI), b/ 3-glycidoxypropyltrimethoxysilane (GPTMS), c/ 3-aminopropyltriethoxysilane (APS). Composition, concentration and pH of several background electrolyte were tested, namely citrate, phosphate, aminocaproate and TRIS. The capillary surface was characterized by determination of the electroosmotic flow (EOF). The conditions were optimized by running a standard mixture of proteins (cytochrome C, lysozyme, ribonuclease A) and then applied to [Gly-14]-humanin.