Sub 2-μm macroporous silica particles derivatized for enhanced lectin affinity enrichment of glycoproteins.
Sub 2-μm macroporous silica particles derivatized for enhanced lectin affinity enrichment of glycoproteins.
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DOI：
10.1021/ac303274w
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发表时间：
2013-02-05
影响因子：
7.4
通讯作者：
Novotny MV
中科院分区：
文献类型：
--
作者：
Mann BF;Mann AK;Skrabalak SE;Novotny MV
一种新的，机械稳定的二氧化硅微粒与大尺寸的内部孔（1.6 μm的颗粒与100 nm的孔）已被开发用于色谱。通过透射电子显微镜（TEM）观察，颗粒的特征在于具有高颗粒内空隙体积的互连大孔的广泛网络。它们是通过称为超声喷雾热解（USP）的气溶胶组装技术合成的。该颗粒具有大孔材料的高表面积，约200 m2/g，使其适用于大生物分子分离。为了证明它们的生物分离潜力，它们已经用凝集素功能化，用于糖蛋白的亲和富集。用两种凝集素，伴刀豆球蛋白A（Con A）和橙黄粉虱凝集素（AAL）衍生化该材料，并用标准糖蛋白测试结合特性。在50 × 1 mm柱中，Con A可与75 μg标准糖蛋白结合。在初始测试之后，凝集素微柱用于从1 μL体积的血清样品中富集糖蛋白，对于每种凝集素进行三次重复。富集的血清级分进行了并行糖组学和糖蛋白质组学分析与质谱，以表明新的颗粒提供了宝贵的生物样品材料的微量分析优良的灵敏度。大孔结构和小粒径的独特组合表明该材料可能具有超高压液相色谱（UHPLC）形式中大分子色谱分离常规模式的优势。
A new, mechanically stable silica microparticle with macro-sized internal pores (1.6-μm particles with 100-nm pores) has been developed for chromatography. The particles are characterized by an extensive network of interconnected macropores with a high intraparticle void volume as observed by transmission electron microscopy (TEM). They are synthesized by an aerosol assembly technique called ultrasonic spray pyrolysis (USP). The particles have a high surface area for a macroporous material, ~200 m2/g, making them suitable for large biomolecular separations. To demonstrate their potential for bioseparations, they have been functionalized with lectins for affinity enrichment of glycoproteins. The material was derivatized with two lectins, Concanavalin A (Con A) and Aleuria aurantia lectin (AAL), and binding properties were tested with standard glycoproteins. The columns exhibited excellent binding capacities for microaffinity enrichment—Con A was able to bind 75 μg of a standard glycoprotein in a 50 × 1-mm column. Following initial tests, the lectin microcolumns were utilized for enrichment of glycoproteins from 1-μL volumes of blood serum samples, which was performed in triplicate for each lectin. The enriched serum fractions were subjected to side-by-side glycomic and glycoproteomic profiling analyses with mass spectrometry to show that the new particles offer excellent sensitivity for microscale analyses of precious biological sample materials. The unique combination of the macroporous architecture and small particle diameter suggests the material may have advantages for conventional modes of chromatographic separation of macromolecules in an ultrahigh-pressure liquid chromatography (UHPLC) format.
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影响因子：
7.4
作者：
Alley, William R., Jr.;Madera, Milan;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
影响因子：
4.4
作者：
Lin, Zhenxin;Simeone, Diane M.;Anderson, Michelle A.;Brand, Randall E.;Xie, Xiaolei;Shedden, Kerby A.;Ruffin, Mack T.;Lubman, David M.
通讯作者：
Lubman, David M.
影响因子：
3.7
作者：
HAPPEL, J
通讯作者：
HAPPEL, J
影响因子：
7.4
作者：
KIRKLAND, JJ
通讯作者：
KIRKLAND, JJ
DOI：
10.1146/annurev.anchem.1.031207.113014
发表时间：
2010-01-01
期刊：
ANNUAL REVIEW OF ANALYTICAL CHEMISTRY, VOL 3
影响因子：
--
作者：
Jorgenson, James W.
通讯作者：
Jorgenson, James W.