Polymer microchip CE of proteins either off- or on-chip labeled with chameleon dye for simplified analysis.

Electrophoresis
Ming YuAdam T Woolley

Abstract

Microchip CE of proteins labeled either off- or on-chip with the "chameleon" CE dye 503 using poly(methyl methacrylate) microchips is presented. A simple dynamic coating using the cationic surfactant CTAB prevented nonspecific adsorption of protein and dye to the channel walls. The labeling reactions for both off- and on-chip labeling proceeded at room temperature without requiring heating steps. In off-chip labeling, a 9 ng/mL concentration detection limit for BSA, corresponding to a approximately 7 fg (100 zmol) mass detection limit, was obtained. In on-chip tagging, the free dye and protein were placed in different reservoirs of the microchip, and an extra incubation step was not needed. A 1 microg/mL concentration detection limit for BSA, corresponding to a approximately 700 fg (10 amol) mass detection limit, was obtained from this protocol. The earlier elution time of the BSA peak in on-chip labeling resulted from fewer total labels on each protein molecule. Our on-chip labeling method is an important part of automation in miniaturized devices.

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Citations

Sep 28, 1998·Biochimica Et Biophysica Acta·T IyanagiS Ikushiro
Sep 22, 2012·Journal of Chromatography. a·Pamela N NgeAdam T Woolley
Jul 23, 2013·Protein Science : a Publication of the Protein Society·Glareh AzadiAnubhav Tripathi
Dec 10, 2014·Electrophoresis·Jayson V PagaduanAdam T Woolley
Sep 13, 2011·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Xiaofang GaoTakehiko Kitamori
May 13, 2017·Analytical Methods : Advancing Methods and Applications·Mukul SonkerAdam T Woolley
May 14, 2010·Analytical Chemistry·Arun AroraAndreas Manz

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