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Electrophoresis


Title:Carboxymethyl chitosan-coated capillary and its application in CE of proteins
Author(s):Fu X; Huang L; Gao F; Li W; Pang N; Zhai M; Liu H; Wu M;
Address:"Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing, PR China"
Journal Title:Electrophoresis
Year:2007
Volume:28
Issue:12
Page Number:1958 - 1963
DOI: 10.1002/elps.200600558
ISSN/ISBN:0173-0835 (Print) 0173-0835 (Linking)
Abstract:"A hydrophilic basic polysaccharide, carboxymethyl chitosan (CMC) as a capillary coating is presented with a simple preparation procedure. The CMC-coated capillary showed a long lifetime of more than 100 runs, and had good tolerance to some organic solvents, 0.1 M HCl, 0.1 M NaOH, and 5 M urea. The run-to-run, day-to-day, and capillary-to-capillary RSDs for the CMC-coated capillary were all below 2.0% for the determination of EOF. Moreover, the coatings with different concentrations and molecular weights of CMC were also investigated. The CMC-coated capillary was successfully applied to separate basic proteins and recombinant human erythropoietin (rhEPO). Furthermore, several experimental parameters, such as the concentration and pH of the running buffer, temperature, and applied voltage, were optimized for the separation of rhEPO glycoforms. Comparison of an uncoated capillary with chitosan- and CMC-coated capillaries for the separation of rhEPO glycoforms was also discussed. The results demonstrated that rhEPO glycoforms can be well separated by a CMC-coated capillary within 8 min with good reproducibility and resolution. Finally, the volatile BGE HAc-NH4Ac was utilized to separate rhEPO for its further application with CE-MS, achieving a satisfactory result"
Keywords:"Adsorption Buffers Chitosan/*analogs & derivatives/*chemistry Coated Materials, Biocompatible/*chemistry Electrophoresis, Capillary/*instrumentation/methods Erythropoietin/analysis Humans Protein Isoforms/analysis Proteins/*analysis Quaternary Ammonium Co;"
Notes:"MedlineFu, Xiaofang Huang, Ling Gao, Fei Li, Wei Pang, Nannan Zhai, Maolin Liu, Huwei Wu, Moutian eng Comparative Study Evaluation Study Research Support, Non-U.S. Gov't Germany 2007/05/10 Electrophoresis. 2007 Jun; 28(12):1958-63. doi: 10.1002/elps.200600558"

 
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