Over 1000-fold protein preconcentration for microliter-volume samples at a pH junction using capillary electrophoresis
| Autor: | Josephine T.-M. Lo, Chandra A. Nesbitt, Ken K.-C. Yeung |
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| Rok vydání: | 2005 |
| Předmět: |
Chromatography
Capillary action Chemistry Organic Chemistry Lipid Bilayers Analytical chemistry Electrophoresis Capillary Proteins General Medicine Hydrogen-Ion Concentration Biochemistry Analytical Chemistry Sample volume chemistry.chemical_compound Capillary electrophoresis Isoelectric point Myoglobin Sample preparation Isoelectric Point Lipid bilayer Phospholipids Protein adsorption |
| Zdroj: | Journal of chromatography. A. 1073(1-2) |
| ISSN: | 0021-9673 |
| Popis: | An effective protein preconcentration technique specifically designed for microliter-volume samples is presented. The preconcentration is based on the capturing of protein in its isoelectric point (pI) within an applied electric field, using a pH junction created by a discontinuous buffer system. The buffers were chosen to selectively preconcentrate proteins of neutral pI, myoglobin in this case, while removing other proteins with acidic or basic pIs. For the suppression of electro-osmotic flow (EOF) and protein adsorption, the capillary inner wall was modified with a zwitterionic phospholipid bilayer coating. A preconcentration factor of up to 1700 was obtained for a 1 microg/mL solution of myoglobin. The preconcentration was completed in approximately 20 min. Several sample introduction conditions were presented to accommodate sample volume from one to a few hundreds of microliters. The final volume of the preconcentrated sample band was estimated to be approximately 5 nL. |
| Databáze: | OpenAIRE |
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