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Moving reaction boundary electrophoresis: concept, method and its use to bioseparation and analytical biochemistry

Moving reaction boundary electrophoresis: concept, method an...

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中国化学会第十八届全国有机分析及生物分析学术研讨会

作者： Cheng-xi Cao Laboratory of Bioseparation and Analytical Biochemistry School of Life Science and BiotechnologyShanghai Jiao Tong University

During the last two decades, the concept of moving reaction boundary（MRB）electrophoresis was developed by our lab, including neutralization boundary（MNB）,precipitate boundary（MPB）, chelation boundary（MCB）, redox boundary（MROB） and affinity boundary（MAB） as well as supramolecular boundary（MSB）. The concept of MRB has been used in the following bioseparations and biochemical ***, a series of online MRB-based sample stacking methods have been developed for the increase of sensitivity of capillary electrophoresis（CE）. The developed methods concerned MNB-, MCB-,MPB-, MAB- and MSB-based sample stacking. The analytes included drugs, zwitterionic solutes, heavy metal ions and ***, we proposed the MRB-based dynamic theory of isoelectric focusing（IEF）,illuminated the mechanism of IEF instability existing in IEF for 50 years and Hjerten’s mobilization of pH gradient, and developed of IEF with high stability, speed and ***, a novel MSB was proposed by using monosaccharide or glycoprotein in microcolumn and the complex probe of BBV-HPTS in chip electrophoresis. The MSB experiments revealed that（i） the probe could interact with free glucose or bound one;（ii）the detection signal was as function of concentration of glucose or glycoprotein;and（iii）the band motion has relation with glucose ***, a novel MRB electrophoretic titration method was developed for acid, base,protein content and structure analyses. In the developed method, the MRB was created between the hydroxide ions in the cathodic vessel and the free acidic groups of protein immobilized via highly cross-linked polyacrylamide gel. It was observed that the boundary velocity（VMRB） was as a function of protein content. The experiments revealed that good linear calibration functions between VMRB and protein content（correlation coefficients R＞0.98）. The experiments further demonstrated the following merits of developed method:（i）weak influence of non-protein nitrogen additive（

关键词： capillary electrophoresis isoelectric focusing moving reaction boundary protein content residue sample stacking and protein titration

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