Use of organic solvent to prevent protein adsorption in CE-MS experiments

被引:38
|
作者
Staub, Aline [1 ,2 ]
Comte, Sylvain [1 ]
Rudaz, Serge [1 ,2 ]
Veuthey, Jean-Luc [1 ,2 ]
Schappler, Julie [1 ,2 ]
机构
[1] Univ Geneva, Sch Pharmaceut Sci, Univ Lausanne, CH-1211 Geneva 4, Switzerland
[2] Univ Geneva, SCAHT, CMU, CH-1211 Geneva 4, Switzerland
关键词
Adsorption; Capillary electrophoresis; Organic solvent; Proteins; CAPILLARY-ELECTROPHORESIS; COATED CAPILLARIES; COATING CONDITIONS; MASS-SPECTROMETRY; SEPARATIONS; EFFICIENCY; STABILITY; SILICA;
D O I
10.1002/elps.201000245
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Protein adsorption onto capillary wall often hampers CE separations, particularly in the CZE mode. Electrostatic interactions are not the only factors affecting adsorption, as hydrophobic interactions and/or protein conformational changes are also involved in the adsorption phenomenon. Numerous methods can be used to reduce or avoid adsorption, such as (i) addition of low molecular weight molecules in the BGE, (ii) use of surfactants, or (iii) capillary coatings. However, most of these methods are not MS-compatible. In this study, we evaluated the addition of organic solvent as an alternative MS-compatible method to decrease protein adsorption. The effect of the solvent addition was emphasized using classical methods for estimating reversible and irreversible adsorption. In many cases, organic solvents were effective at decreasing adsorption. However, the influence of the organic solvent on protein adsorption should be evaluated case-by-case in CE method development.
引用
收藏
页码:3326 / 3333
页数:8
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