Mobility-based wall adsorption isotherms for comparing capillary electrophoresis with single-molecule observations.
The adsorption properties of R-phycoerythrin (RPE), an autofluorescent protein, on the fused-silica surface were studied in capillary electrophoresis (CE) and in single-molecule experiments. The band shapes and migration times were measured in CE, and adsorption and desorption events were recorded at the single-molecule level by imaging within the evanescent field layer using total internal reflection fluorescence microscopy. The adsorbed RPE molecules on the surface of the fused-silica prism were counted with confidence based on ImageJ software. The capacity factor and desorption rate were estimated from the counting results. The mobility-based adsorption isotherms were constructed from both computer simulations and experiments to determine the capacity factor.
Fang
Ning
N
Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
Zhang
Hui
H
Li
Jiangwei
J
Li
Hung-Wing
HW
Yeung
Edward S
ES
D000327:Adsorption; D003198:Computer Simulation; D019075:Electrophoresis, Capillary; D007700:Kinetics; D008856:Microscopy, Fluorescence; D010799:Phycoerythrin; D012984:Software
D010799:Phycoerythrin