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Open-tubular capillary electrochromatography of bovine β-lactoglobulin variants A and B using an aptamer stationary phase
被引:1
作者:
Rehder, MA
[1
]
McGown, LB
[1
]
机构:
[1] Duke Univ, PM Gross Chem Lab, Dept Chem, Durham, NC 27705 USA
关键词:
aptamers;
capillary electrochromatography;
open-tubular capillary electrochromatography;
protein separation;
beta-Lactoglobulin;
D O I:
10.1002/1522-2683(200109)22:17<3759::AID-ELPS3759>3.0.CO;2-C
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
DNA aptamers that form a G-quartet conformation were covalently attached to a capillary surface for open-tubular capillary electrochromatographic separation of bovine beta -lactoglobulin variants A and B, which vary by 2 of their 162 amino acid residues. Separation was achieved using a 4-plane, G-quartet aptamer stationary phase with tris(hydroxymethyl)aminomethane (Tris) or phosphate buffer as the mobile phase. In control experiments, separation did not occur using either an oligonucleotide of similar base composition but which does not form a G-quartet structure, or using capillary zone electrophoresis on a bare capillary under similar experimental conditions. Separation was achieved using a capillary coated only with the covalent linker molecule. In phosphate buffer, the separations were similar for aptamer-coated and linker-only stationary phases, while in Tris buffer, retention times were almost doubled for the linker-only capillary. When Tris buffer is the mobile phase, there appears to be weaker interactions between the proteins and the stationary phase that may result in a gentler, less denaturing separation than is commonly achieved using hydrocarbon-based stationary phases.
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页码:3759 / 3764
页数:6
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