Capillary Electrophoresis / Electrospray Mass Spectrometry Analysis of Oligonucleotides and Native Proteins Using Poly(ethylene oxide) Solution
Date Issued
2011
Date
2011
Author(s)
Chen, Szu-Fang
Abstract
Analyzing oligonucleotides and native proteins by capillary electrophoresis usually adds polymer in the background electrolyte. By using the polymer solution, analytes with similar mobilities can be separated via molecular sieving mechanism. However, PEO will suppress the analyte signal and contaminate the ESI ion source. To alleviate these problems resulting from the introduction of polymers into ESI source, a liquid junction-low flow interface was utilized to hyphenate CE with polymer solution and ESI-MS in the analysis of oligonucleotides and native proteins. In the analysis of negative charged oligonucleotides, polymers would migrate toward the inlet reservoir with EOF under reversed polarity to avoid the introduction of polymers into ion source. In addition, filling the liquid junction with polymer solution could make polymers in the liquid junction reservoir flow into separation column. Thus, the separation efficiency could be maintained. Because of the mobility of analyte is greater than EOF, so it could migrate to MS inlet and be successfully detected. On the other hand, polymers would migrate to MS inlet with EOF under normal polarity in the analysis of intact proteins. As a consequence, a PVA coated capillary column was utilized as the connecting column to make polymer stay in the liquid junction. By this method, the introduction of polymers into ion source could be prevented. And the analyte could migrate to the detection end and be detected by its own mobility.
Subjects
oligonucleotides
polymer solution
PEO
sieving mechanism
CE-MS
native protein
liquid-junction/low flow interface
Type
thesis
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