RESUMEN
The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. In this study, we used the charge characteristics of protein to develop a simple and rapid electro-microfluidic desalting system as an effective means of cleaning up protein sample. When we loaded a urea-rich protein sample and a buffer solution into a free-flow zone electrophoresis (FFZE) chamber, the microfluidic device was able to separate the charged protein sample and the non-charged urea. With a 90 V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors.
Asunto(s)
Técnicas Analíticas Microfluídicas/instrumentación , Microfluídica/métodos , Proteínas/análisis , Animales , Técnicas Biosensibles , Electroforesis/instrumentación , Electroforesis/métodos , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Técnicas Analíticas Microfluídicas/métodos , Microfluídica/instrumentación , Proteínas/química , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Urea/químicaRESUMEN
By using homochiral mesoporous POST-1 (Scheme 1) as an inherently chiral sensitizing host, supramolecular enantiodifferentiating photoisomerizations of (Z)-cyclooctene (1Z) were performed for the first time to examine the sensitizing ability of the chiral nanopores, walls of which are decorated with pyridine chromophore. The homochiral nanopores of POST-1 crystals were demonstrated to function as an effective chiral-sensitizing cavity to give the optically active (E)-isomer 1E in up to 5% enantiomeric excess (ee), thus expanding the scope of porous material-mediated chirogenesis to both ground and excited electronic states.
