Lysine-based zwitterionic molecular micelle for simultaneous separation of acidic and basic proteins using open tubular capillary electrochromatography.
Lysine-based zwitterionic molecular micelle for simultaneous separation of acidic and basic proteins using open tubular capillary electrochromatography.
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DOI：
10.1021/ac902723n
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发表时间：
2010-05-15
影响因子：
7.4
通讯作者：
Warner, Isiah M.
中科院分区：
文献类型：
--
作者：
Moore, Leonard, Jr.;LeJeune, Zorabel M.;Luces, Candace A.;Gates, Arther T.;Li, Min;El-Zahab, Bilal;Garno, Jayne C.;Warner, Isiah M.
本工作合成了一种两性离子分子胶束聚-ε-十一酰赖氨酸钠（poly-ε-SUK），并将其用作开管毛细管电色谱（OT-CEC）中蛋白质分离的涂层。含有羧酸和胺基团的两性离子聚-ε-SUK可以根据背景电解质的pH而被质子化或去质子化;因此，可以实现总体带正电荷或带负电荷的涂层。该两性离子涂层允许根据背景电解质的pH以正极性或反极性模式分离蛋白质。蛋白质混合物含有4种碱性蛋白（溶菌酶、细胞色素c、α-胰凝乳蛋白酶原A和核糖核酸酶A）和6种酸性蛋白（肌红蛋白、β-乳球蛋白A、β-乳球蛋白B、α-乳白蛋白和白蛋白）。针对酸性（反向模式）和碱性（正常模式）pH值专门优化蛋白质分离。通过改变聚合物和盐浓度来改变聚合物厚度对蛋白质分离度有很大影响，而使用0.4%的优化聚-ε-SUK涂层浓度，在两种模式下所有峰也基线分离。在优化的酸性和碱性条件下分离人血清中的蛋白质，以证明该涂层的一般效用。利用原子力显微镜（AFM）对硅表面的聚-ε-SUK胶束涂层进行了纳米级表征，以深入了解两性离子涂层的形貌和厚度。聚合物涂层的厚度范围为0.9-2.9 nm，基于使用nanoshaving的局部测量，nanoshaving是一种基于AFM的纳米光刻方法。
In this work, a zwitterionic molecular micelle, poly-ε-sodium-undecanoyl lysinate (poly-ε-SUK), was synthesized and employed as a coating in open tubular capillary electrochromatography (OT-CEC) for protein separation. The zwitterionic poly-ε-SUK containing both carboxylic acid and amine groups can be either protonated or deprotonated depending on the pH of the background electrolyte; therefore, either an overall positively or negatively charged coating can be achieved. This zwitterionic coating allows protein separations in either normal or reverse polarity mode depending on the pH of the background electrolyte. The protein mixtures contained 4 basic proteins (lysozyme, cytochrome c, α-chymotrypsinogen A, and ribonuclease A) and 6 acidic proteins (myoglobin, β-lactoglobulin A, β-lactoglobulin B, α-lactalbumin, and albumin). Protein separations were optimized specifically for acidic (reverse mode) and basic (normal mode) pH values. Varying the polymer thickness by changing the polymer and salt concentration had a great influence on protein resolution, while all peaks were also baseline resolved in both modes using the optimized poly-ε-SUK coating concentration of 0.4%. Proteins in human sera were separated under optimized acidic and basic conditions in order to demonstrate the general utility of this coating. Nanoscale characterizations of the poly-ε-SUK micellar coatings on silicon surfaces were accomplished using atomic force microscopy (AFM), to gain insight into the morphology and thickness of the zwitterionic coating. The thickness of the polymer coating ranged from 0.9–2.9 nm based on local measurements using nanoshaving, an AFM-based method of nanolithography.
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影响因子：
7.4
作者：
Graul, TW;Schlenoff, JB
通讯作者：
Schlenoff, JB
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作者：
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